Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 1.332
Filtrar
1.
Nat Commun ; 11(1): 1092, 2020 02 27.
Artigo em Inglês | MEDLINE | ID: mdl-32107390

RESUMO

Micro(mi)RNA-based post-transcriptional regulatory mechanisms have been broadly implicated in the assembly and modulation of synaptic connections required to shape neural circuits, however, relatively few specific miRNAs have been identified that control synapse formation. Using a conditional transgenic toolkit for competitive inhibition of miRNA function in Drosophila, we performed an unbiased screen for novel regulators of synapse morphogenesis at the larval neuromuscular junction (NMJ). From a set of ten new validated regulators of NMJ growth, we discovered that miR-34 mutants display synaptic phenotypes and cell type-specific functions suggesting distinct downstream mechanisms in the presynaptic and postsynaptic compartments. A search for conserved downstream targets for miR-34 identified the junctional receptor CNTNAP4/Neurexin-IV (Nrx-IV) and the membrane cytoskeletal effector Adducin/Hu-li tai shao (Hts) as proteins whose synaptic expression is restricted by miR-34. Manipulation of miR-34, Nrx-IV or Hts-M function in motor neurons or muscle supports a model where presynaptic miR-34 inhibits Nrx-IV to influence active zone formation, whereas, postsynaptic miR-34 inhibits Hts to regulate the initiation of bouton formation from presynaptic terminals.


Assuntos
Proteínas de Ligação a Calmodulina/genética , Moléculas de Adesão Celular Neuronais/genética , Proteínas de Drosophila/genética , Regulação da Expressão Gênica no Desenvolvimento , MicroRNAs/metabolismo , Terminações Pré-Sinápticas/fisiologia , Animais , Animais Geneticamente Modificados , Proteínas de Ligação a Calmodulina/metabolismo , Moléculas de Adesão Celular Neuronais/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/fisiologia , Larva/crescimento & desenvolvimento , Morfogênese/genética , Mutação , Junção Neuromuscular/citologia , Junção Neuromuscular/crescimento & desenvolvimento
2.
BMC Plant Biol ; 20(1): 38, 2020 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-31992205

RESUMO

BACKGROUD: Many Gram-negative bacteria use N-acyl-homoserine lactones (AHLs) to communicate each other and to coordinate their collective behaviors. Recently, accumulating evidence shows that host plants are able to sense and respond to bacterial AHLs. Once primed, plants are in an altered state that enables plant cells to more quickly and/or strongly respond to subsequent pathogen infection or abiotic stress. RESULTS: In this study, we report that pretreatment with N-3-oxo-octanoyl-homoserine lactone (3OC8-HSL) confers resistance against the pathogenic bacterium Pseudomonas syringae pv. tomato DC3000 (PstDC3000) in Arabidopsis. Pretreatment with 3OC8-HSL and subsequent pathogen invasion triggered an augmented burst of hydrogen peroxide, salicylic acid accumulation, and fortified expression of the pathogenesis-related genes PR1 and PR5. Upon PstDC3000 challenge, plants treated with 3OC8-HSL showed increased activities of defense-related enzymes including peroxidase, catalase, phenylalanine ammonialyase, and superoxide dismutase. In addition, the 3OC8-HSL-primed resistance to PstDC3000 in wild-type plants was impaired in plants expressing the bacterial NahG gene and in the npr1 mutant. Moreover, the expression levels of isochorismate synthases (ICS1), a critical salicylic acid biosynthesis enzyme, and two regulators of its expression, SARD1 and CBP60g, were potentiated by 3OC8-HSL pretreatment followed by pathogen inoculation. CONCLUSIONS: Our data indicate that 3OC8-HSL primes the Arabidopsis defense response upon hemibiotrophic bacterial infection and that 3OC8-HSL-primed resistance is dependent on the SA signaling pathway. These findings may help establish a novel strategy for the control of plant disease.


Assuntos
4-Butirolactona/análogos & derivados , Arabidopsis , Imunidade Vegetal/efeitos dos fármacos , Pseudomonas syringae/patogenicidade , Ácido Salicílico/metabolismo , 4-Butirolactona/farmacologia , Arabidopsis/genética , Arabidopsis/microbiologia , Proteínas de Arabidopsis/efeitos dos fármacos , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Ligação a Calmodulina/efeitos dos fármacos , Proteínas de Ligação a Calmodulina/genética , Proteínas de Ligação a Calmodulina/metabolismo , Genes de Plantas , Transferases Intramoleculares/efeitos dos fármacos , Transferases Intramoleculares/genética , Transferases Intramoleculares/metabolismo , Doenças das Plantas/microbiologia , Imunidade Vegetal/genética , Percepção de Quorum/fisiologia , Transdução de Sinais/efeitos dos fármacos
3.
Nucleic Acids Res ; 48(3): 1192-1205, 2020 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-31950163

RESUMO

Somatic synonymous mutations are one of the most frequent genetic variants occurring in the coding region of cancer genomes, while their contributions to cancer development remain largely unknown. To assess whether synonymous mutations involved in post-transcriptional regulation contribute to the genetic etiology of cancers, we collected whole exome data from 8,320 patients across 22 cancer types. By employing our developed algorithm, PIVar, we identified a total of 22,948 posttranscriptionally impaired synonymous SNVs (pisSNVs) spanning 2,042 genes. In addition, 35 RNA binding proteins impacted by these identified pisSNVs were significantly enriched. Remarkably, we discovered markedly elevated ratio of somatic pisSNVs across all 22 cancer types, and a high pisSNV ratio was associated with worse patient survival in five cancer types. Intriguing, several well-established cancer genes, including PTEN, RB1 and PIK3CA, appeared to contribute to tumorigenesis at both protein function and posttranscriptional regulation levels, whereas some pisSNV-hosted genes, including UBR4, EP400 and INTS1, exerted their function during carcinogenesis mainly via posttranscriptional mechanisms. Moreover, we predicted three drugs associated with two pisSNVs, and numerous compounds associated with expression signature of pisSNV-hosted genes. Our study reveals the prevalence and clinical relevance of pisSNVs in cancers, and emphasizes the importance of considering posttranscriptional impaired synonymous mutations in cancer biology.


Assuntos
Carcinogênese/genética , Genoma Humano/genética , Neoplasias/genética , Mutação Silenciosa/genética , Adulto , Idoso , Proteínas de Ligação a Calmodulina/genética , Classe I de Fosfatidilinositol 3-Quinases/genética , DNA Helicases/genética , Proteínas de Ligação a DNA/genética , Exoma/genética , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Neoplasias/classificação , Neoplasias/patologia , PTEN Fosfo-Hidrolase/genética , Intervalo Livre de Progressão , Processamento de Proteína Pós-Traducional/genética , Locos de Características Quantitativas/genética , Proteínas de Ligação a Retinoblastoma/genética , Ubiquitina-Proteína Ligases/genética , Proteína Wnt1/genética
4.
Plant Physiol ; 181(3): 1314-1327, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31548265

RESUMO

Calmodulin (CaM) regulates plant disease responses through its downstream calmodulin-binding proteins (CaMBPs) often by affecting the biosynthesis or signaling of phytohormones, such as jasmonic acid (JA) and salicylic acid. However, how these CaMBPs mediate plant hormones and other stress resistance-related signaling remains largely unknown. In this study, we conducted analyses in Arabidopsis (Arabidopsis thaliana) on the functions of AtIQM1 (IQ-Motif Containing Protein1), a Ca2+-independent CaMBP, in JA biosynthesis and defense against the necrotrophic pathogen Botrytis cinerea using molecular, biochemical, and genetic analyses. IQM1 directly interacted with and promoted CATALASE2 (CAT2) expression and CAT2 enzyme activity and indirectly increased the activity of the JA biosynthetic enzymes ACX2 and ACX3 through CAT2, thereby positively regulating JA content and B. cinerea resistance. In addition, in vitro assays showed that in the presence of CaM5, IQM1 further enhanced the activity of CAT2, suggesting that CaM5 may affect the activity of CAT2 by combining with IQM1 in the absence of Ca2+ Our data indicate that IQM1 is a key regulatory factor in signaling of plant disease responses mediated by JA. The study also provides new insights that CaMBP may play a critical role in the cross talk of multiple signaling pathways in the context of plant defense processes.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Botrytis/fisiologia , Proteínas de Ligação a Calmodulina/metabolismo , Doenças das Plantas/imunologia , Reguladores de Crescimento de Planta/metabolismo , Motivos de Aminoácidos , Arabidopsis/enzimologia , Arabidopsis/imunologia , Arabidopsis/microbiologia , Proteínas de Arabidopsis/genética , Sinalização do Cálcio , Proteínas de Ligação a Calmodulina/genética , Ciclopentanos/metabolismo , Resistência à Doença , Oxilipinas/metabolismo , Doenças das Plantas/microbiologia , Estômatos de Plantas/enzimologia , Estômatos de Plantas/genética , Estômatos de Plantas/imunologia , Estômatos de Plantas/microbiologia , Ácido Salicílico/metabolismo
5.
Cell Struct Funct ; 44(2): 95-104, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31548446

RESUMO

KCBP is a microtubule (MT) minus-end-directed kinesin widely conserved in plants. It was shown in Arabidopsis that KCBP controls trichome cell shape by orchestrating MT and actin cytoskeletons using its tail and motor domains. In contrast, the KCBP knockout (KO) line in the moss Physcomitrella patens showed a defect in nuclear and organelle positioning in apical stem cells. Moss KCBP is postulated to transport the nucleus and chloroplast via direct binding to their membranes, since it binds to and transports liposomes composed of phospholipids in vitro. However, domains required for cargo transport in vivo have not been mapped. Here, we performed a structure-function analysis of moss KCBP. We found that the FERM domain in the tail region, which is known to bind to lipids as well as other proteins, is essential for both nuclear and chloroplast positioning, whereas the proximal MyTH4 domain plays a supporting role in chloroplast transport. After anaphase but prior to nuclear envelope re-formation, KCBP accumulates on the chromosomes, in particular at the centromeric region in a FERM-dependent manner. In the KCBP KO line, the rate of poleward chromosome movement in anaphase was reduced and lagging chromosomes occasionally appeared. These results suggest that KCBP binds to non-membranous naked chromosomes via an unidentified protein(s) for their transport. Finally, the liverwort orthologue of KCBP rescued the chromosome/chloroplast mis-positioning of the moss KCBP KO line, suggesting that the cargo transport function is conserved at least in bryophytes.Key words: kinesin, mitosis, chromosome segregation, kinetochore, dynein.


Assuntos
Anáfase , Proteínas de Arabidopsis/metabolismo , Arabidopsis/citologia , Arabidopsis/metabolismo , Proteínas de Ligação a Calmodulina/metabolismo , Cromátides/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Ligação a Calmodulina/deficiência , Proteínas de Ligação a Calmodulina/genética
6.
Life Sci Alliance ; 2(3)2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31167803

RESUMO

WT1 is a transcriptional activator that controls the boundary between multipotency and differentiation. The transcriptional cofactor BASP1 binds to WT1, forming a transcriptional repressor complex that drives differentiation in cultured cells; however, this proposed mechanism has not been demonstrated in vivo. We used the peripheral taste system as a model to determine how BASP1 regulates the function of WT1. During development, WT1 is highly expressed in the developing taste cells while BASP1 is absent. By the end of development, BASP1 and WT1 are co-expressed in taste cells, where they both occupy the promoter of WT1 target genes. Using a conditional BASP1 mouse, we demonstrate that BASP1 is critical to maintain the differentiated state of adult taste cells and that loss of BASP1 expression significantly alters the composition and function of these cells. This includes the de-repression of WT1-dependent target genes from the Wnt and Shh pathways that are normally only transcriptionally activated by WT1 in the undifferentiated taste cells. Our results uncover a central role for the WT1-BASP1 complex in maintaining cell differentiation in vivo.


Assuntos
Proteínas de Ligação a Calmodulina/metabolismo , Diferenciação Celular , Proteínas do Citoesqueleto/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Papilas Gustativas/citologia , Papilas Gustativas/metabolismo , Proteínas WT1/metabolismo , Animais , Biomarcadores , Proteínas de Ligação a Calmodulina/genética , Diferenciação Celular/genética , Proteínas do Citoesqueleto/genética , Imunofluorescência , Expressão Gênica , Técnicas de Silenciamento de Genes , Camundongos , Camundongos Transgênicos , Modelos Biológicos , Proteínas do Tecido Nervoso/genética , Fenótipo , Ligação Proteica , Proteínas WT1/genética
7.
Artigo em Inglês | MEDLINE | ID: mdl-31202182

RESUMO

The multifactorial nature of Late Onset Alzheimer's Disease (LOAD), the AD form of major relevance on epidemiological and social aspects, has driven the original investigation by LC-MS and top-down proteomics approach of the protein repertoire of the brain tissue of TgCRND8 model mice fed with a diet deficient in B vitamins. The analysis of the acid-soluble fraction of brain tissue homogenates identified a list of proteins and peptides, proteoforms and PTMs. In order to disclose possible modulations, their relative quantification in wild type and AD model mice under both B vitamin deficient and control diets was performed. The levels of metallothionein III, guanine nucleotide-binding protein G(I)/G(S)/G(O) subunit gamma-2 and brain acid soluble protein 1 showed statistically significant alterations depending on genotype, diet or both effects, respectively. Particularly, metallothionein III exhibited increased levels in TgCRND8 mice under B vitamin deficient diet with respect to wild type mice under both diets. Brain acid soluble protein 1 showed the opposite, revealing decreased levels in all diet groups of AD model mice with respect to wild type mice in control diet. Lower levels of brain acid soluble protein 1 were also observed in wild type mice under deficiency of B vitamins. These results, besides contributing to increase the knowledge of AD at molecular level, give new suggestions for deeply investigating metallothionein III and brain acid soluble protein 1 in AD.


Assuntos
Doença de Alzheimer/metabolismo , Encéfalo/metabolismo , Hiper-Homocisteinemia/metabolismo , Proteoma/metabolismo , Complexo Vitamínico B/análise , Doença de Alzheimer/genética , Peptídeos beta-Amiloides/metabolismo , Animais , Química Encefálica , Proteínas de Ligação a Calmodulina/genética , Proteínas de Ligação a Calmodulina/metabolismo , Cromatografia Líquida , Proteínas do Citoesqueleto/genética , Proteínas do Citoesqueleto/metabolismo , Modelos Animais de Doenças , Feminino , Humanos , Hiper-Homocisteinemia/etiologia , Hiper-Homocisteinemia/genética , Masculino , Espectrometria de Massas , Camundongos , Camundongos Transgênicos , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Proteoma/química , Proteoma/genética , Complexo Vitamínico B/metabolismo
8.
Invest Ophthalmol Vis Sci ; 60(7): 2551-2562, 2019 06 03.
Artigo em Inglês | MEDLINE | ID: mdl-31212307

RESUMO

Purpose: To use supervised machine learning to predict visual function from retinal structure in retinitis pigmentosa (RP) and apply these estimates to CEP290- and NPHP5-associated Leber congenital amaurosis (LCA) to determine the potential for functional improvement. Methods: Patients with RP (n = 20) and LCA due to CEP290 (n = 12) or NPHP5 (n = 6) mutations were studied. A patient with CEP290 mutations but mild retinal degeneration was included. RP patients had cone-mediated macular function. A machine learning technique was used to associate perimetric sensitivities to local structure in RP patients. Models trained on RP data were applied to predict visual function in LCA. Results: The RP and LCA patients had comparable retinal structure. RP patients had peak sensitivity at the fovea surrounded by decreasing sensitivity. Machine learning could successfully predict perimetry results from segmented or unsegmented optical coherence tomography (OCT) input. Application of machine learning predictions to LCA within the residual macular island of photoreceptor structure showed differences between predicted and measured sensitivities defining treatment potential. In patients with retained vision, the treatment potential was 4.6 ± 2.9 dB at the fovea but 16.4 ± 4.4 dB at the parafovea. In patients with limited or no vision, the treatment potential was 17.6 ± 9.4 dB. Conclusions: Cone vision improvement potential in LCA due to CEP290 or NPHP5 mutations is predictable from retinal structure using a machine learning approach. This should allow individual prediction of the maximal efficacy in clinical trials and guide decisions about dosing. Similar strategies can be used in other retinal degenerations to estimate the extent and location of treatment potential.


Assuntos
Antígenos de Neoplasias/genética , Proteínas de Ligação a Calmodulina/genética , Defeitos da Visão Cromática/terapia , Amaurose Congênita de Leber/genética , Amaurose Congênita de Leber/terapia , Aprendizado de Máquina , Proteínas de Neoplasias/genética , Células Fotorreceptoras Retinianas Cones/fisiologia , Adolescente , Adulto , Defeitos da Visão Cromática/genética , Feminino , Terapia Genética , Humanos , Amaurose Congênita de Leber/fisiopatologia , Masculino , Pessoa de Meia-Idade , Mutação , Retinite Pigmentosa/genética , Retinite Pigmentosa/fisiopatologia , Retinite Pigmentosa/terapia , Tomografia de Coerência Óptica/métodos , Testes de Campo Visual , Campos Visuais/fisiologia , Adulto Jovem
9.
Plant Mol Biol ; 101(1-2): 21-40, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31049793

RESUMO

KEY MESSAGE: Arabidopsis thaliana mlo3 mutant plants are not affected in pathogen infection phenotypes but-reminiscent of mlo2 mutant plants-exhibit spontaneous callose deposition and signs of early leaf senescence. The family of Mildew resistance Locus O (MLO) proteins is best known for its profound effect on the outcome of powdery mildew infections: when the appropriate MLO protein is absent, the plant is fully resistant to otherwise virulent powdery mildew fungi. However, most members of the MLO protein family remain functionally unexplored. Here, we investigate Arabidopsis thaliana MLO3, the closest relative of AtMLO2, AtMLO6 and AtMLO12, which are the Arabidopsis MLO genes implicated in the powdery mildew interaction. The co-expression network of AtMLO3 suggests association of the gene with plant defense-related processes such as salicylic acid homeostasis. Our extensive analysis shows that mlo3 mutants are unaffected regarding their infection phenotype upon challenge with the powdery mildew fungi Golovinomyces orontii and Erysiphe pisi, the oomycete Hyaloperonospora arabidopsidis, and the bacterial pathogen Pseudomonas syringae (the latter both in terms of basal and systemic acquired resistance), indicating that the protein does not play a major role in the response to any of these pathogens. However, mlo3 genotypes display spontaneous callose deposition as well as signs of early senescence in 6- or 7-week-old rosette leaves in the absence of any pathogen challenge, a phenotype that is reminiscent of mlo2 mutant plants. We hypothesize that de-regulated callose deposition in mlo3 genotypes might be the result of a subtle transient aberration of salicylic acid-jasmonic acid homeostasis during development.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Proteínas de Ligação a Calmodulina/metabolismo , Resistência à Doença/genética , Glucanos/metabolismo , Doenças das Plantas/imunologia , Arabidopsis/fisiologia , Proteínas de Arabidopsis/genética , Ascomicetos/fisiologia , Proteínas de Ligação a Calmodulina/genética , Ciclopentanos/metabolismo , Genótipo , Homeostase , Mutação , Oomicetos/fisiologia , Oxilipinas/metabolismo , Fenótipo , Doenças das Plantas/microbiologia , Reguladores de Crescimento de Planta/metabolismo , Folhas de Planta/genética , Folhas de Planta/fisiologia , Pseudomonas syringae/fisiologia , Ácido Salicílico/metabolismo
10.
Technol Cancer Res Treat ; 18: 1533033819841433, 2019 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-30947633

RESUMO

Colorectal cancer is one of the most prevalent malignancies worldwide. ENKUR is a transient receptor potential canonical-binding protein that acts as a potential regulator or effector of transient receptor potential canonical channels. It also directly interacts with the p85 regulatory subunit of phosphoinositide 3-kinase. However, the role of ENKUR in colorectal cancer remains unclear. In the present study, the expression profiles of ENKUR in the The Cancer Genome Atlas and ONCOMINE databases were analyzed. Significant downregulation of ENKUR was observed in clinical tumor samples of various cancer types, including colorectal cancer. Decreased ENKUR messenger RNA expression and ENKUR protein level were detected in 6 human colorectal cancer cell lines. Silencing of ENKUR in colorectal cancer cells led to enhanced cell proliferation, migration, and invasion, while the opposite effects were achieved in ENKUR-overexpressing cells. Furthermore, ENKUR-underexpressing cells exhibited increased activity of phosphoinositide 3-kinase /Akt signaling pathway. Downregulation of the epithelial marker, E-cadherin, and upregulation of the mesenchymal markers, vimentin and N-cadherin, were detected in ENKUR-underexpressing cells, suggesting the induction of epithelial-mesenchymal transition. In conclusion, the present study demonstrates that ENKUR may be responsible for alterations in the proliferative, migratory, and invasive potential of colorectal cancer cells through possible involvement in the phosphoinositide 3-kinase /Akt signaling pathway.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Adenocarcinoma/patologia , Proteínas de Ligação a Calmodulina/metabolismo , Neoplasias Colorretais/patologia , Regulação Neoplásica da Expressão Gênica , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/genética , Adenocarcinoma/genética , Adenocarcinoma/metabolismo , Proteínas de Ligação a Calmodulina/genética , Movimento Celular , Proliferação de Células , Neoplasias Colorretais/genética , Neoplasias Colorretais/metabolismo , Transição Epitelial-Mesenquimal , Humanos , Fosfatidilinositol 3-Quinases/genética , Proteínas Proto-Oncogênicas c-akt/genética , Transdução de Sinais , Células Tumorais Cultivadas
11.
Mol Biol Rep ; 46(3): 2721-2732, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30843175

RESUMO

The calmodulin-binding transcriptional activator (CAMTA) family was first observed in tobacco (NtER1) during a screening for the CaM-binding proteins, which are known to be one of the fast response stress proteins. Due to the increased importance of plant transcription factors in recent years; genome-wide identification of CAMTA genes has been performed in several plant species, except for Phaseolus vulgaris. Therefore, our aim was to identify and characterize CAMTA genes in P. vulgaris via in silico genome-wide analysis approach. Our results showed a total of eight CAMTA genes that were identified and observed on five out of 11 chromosomes of P. vulgaris. Four gene couples were found to be segmentally-duplicated and these segmental duplication events were shown to occur from 29.97 to 92.06 MYA. The phylogenetic tree of CAMTA homologs from P. vulgaris, A. thaliana, and G. max. revealed three groups based on their homology and the intron numbers of Pvul-CAMTA genes, ranged from 11 to 12. According to the syteny analysis; CAMTA genes of P. vulgaris and G. max revealed higher similarity, because they have highly similar genomes compared to A. thaliana. All Pvul-CAMTA genes were targeted by miRNAs, which play a role in response mechanism of salt stress. To detect expression levels in different plant tissues, mRNA analysis of Pvul-CAMTA genes were performed using publicly available expression data in Phytozome v12.1. In addition, responses of Pvul-CAMTA genes to salt stress, were also examined via both RNAseq and qRT-PCR analysis. To identify and to obtain insight into biological functions of CAMTA genes in the genome of P. vulgaris, several analyses were conducted using many online and offline bioinformatic tools, genome databases and qRT-PCR analyses. Due to this study being the first in the identification of CAMTA genes in P. vulgaris, this study could be considered as an useful source for future CAMTA genes studies in either P. vulgaris or comparative different plant species.


Assuntos
Proteínas de Ligação a Calmodulina/genética , Phaseolus/genética , Estresse Salino/genética , Mapeamento Cromossômico/métodos , Cromossomos de Plantas/genética , Biologia Computacional , Simulação por Computador , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica de Plantas/genética , Genoma de Planta/genética , Estudo de Associação Genômica Ampla/métodos , Phaseolus/metabolismo , Filogenia , Proteínas de Plantas/genética , Estresse Fisiológico/genética , Transativadores/genética , Ativação Transcricional/genética
12.
Plant Mol Biol ; 99(6): 603-620, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30783953

RESUMO

KEY MESSAGE: Thirty-five IQD genes were identified and analysed in Chinese cabbage and BrIQD5 transgenic plants enhanced the drought resistance of plants. The IQD (IQ67-domain) family plays an important role in various abiotic stress responses in plant species. However, the roles of IQD genes in the Chinese cabbage response to abiotic stress remain unclear. Here, 35 IQD genes, from BrIQD1 to BrIQD35, were identified in Chinese cabbage (Brassica rapa ssp. pekinensis). Based on the phylogenetic analysis, these genes were clustered into three subfamilies (I-III), and members within the same subfamilies shared conserved exon-intron distribution and motif composition. The 35 BrIQD genes were unevenly distributed on 9 of the 10 chromosomes with 4 segmental duplication events. Ka/Ks ratios showed that the duplicated BrIQDs had mainly experienced strong purifying selection. Quantitative real-time polymerase chain reaction of 35 BrIQDs under PEG6000 indicated that BrIQD5 was significantly induced by PEG6000. To verify BrIQD5 function, BrIQD5 was heterologously overexpressed in tobacco and was silenced in Chinese cabbage. BrIQD5-overexpressed plants showed more tolerance to drought stress than wild-type plants, while BrIQD5-silenced plants in Chinese cabbage showed decreased drought tolerance. Additionally, six BrIQD5 potential interactive proteins were isolated by the yeast two-hybrid assay, including BrCaMa, BrCaMb and four other stress-related proteins. Motif IQ1 of BrIQD5 is important for the interaction with BrCaMa and BrCaMb, and the isoleucine in motif IQ1 is an essential amino acid for calmodulin binding to BrIQD5. The identification and cloning of the new Chinese cabbage drought tolerance genes will promote the drought-resistant breeding of Chinese cabbage and help to better understand the mechanism of IQD involved in the drought tolerance of plants.


Assuntos
Proteínas de Arabidopsis/classificação , Proteínas de Arabidopsis/genética , Brassica rapa/genética , Proteínas de Ligação a Calmodulina/classificação , Proteínas de Ligação a Calmodulina/genética , Secas , Regulação da Expressão Gênica de Plantas , Família Multigênica/genética , Filogenia , Mapeamento Cromossômico , Cromossomos de Plantas/genética , Perfilação da Expressão Gênica , Genoma de Planta , Duplicações Segmentares Genômicas , Alinhamento de Sequência , Estresse Fisiológico/genética , Tabaco/genética , Transcriptoma/genética
13.
Pathol Int ; 69(3): 172-176, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30737997

RESUMO

Recently, a new entity "myoepithelioma-like tumor of the vulvar region (MELTVR)" was proposed as a rare mesenchymal neoplasm arising in vulvar regions of adult women. While MELTVRs morphologically resemble soft tissue myoepitheliomas and extraskeletal myxoid chondrosarcomas, they have a unique immunohistochemical profile (positive for epithelial membrane antigen and estrogen receptor, negative for S100 protein and glial fibrillary acidic protein, and loss of INI1/SMARCB1 expression), and lack EWSR1 and NR4A3 gene rearrangement, as seen by fluorescence in situ hybridization. MELTVRs are usually well-demarcated tumors, with no reports of extensive infiltrative growth. In the current report, we present an unusual case of MELTVR showing infiltrative growth and harboring only a few estrogen receptor-positive cells, which might indicate a variation in this rare tumor.


Assuntos
Biomarcadores Tumorais/genética , Rearranjo Gênico/genética , Mioepitelioma/patologia , Receptores Estrogênicos/metabolismo , Proteínas de Ligação a Calmodulina/genética , Humanos , Imuno-Histoquímica/métodos , Mucina-1/imunologia , Mioepitelioma/diagnóstico
14.
New Phytol ; 222(1): 335-348, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30372534

RESUMO

Plants have evolved an array of responses that provide them with protection from attack by microorganisms and other predators. Many of these mechanisms depend upon interactions between the plant hormones jasmonate (JA) and ethylene (ET). However, the molecular basis of these interactions is insufficiently understood. Gene expression and physiological assays with mutants were performed to investigate the role of Arabidopsis BIG gene in stress responses. BIG transcription is downregulated by methyl JA (MeJA), necrotrophic infection or mechanical injury. BIG deficiency promotes JA-dependent gene induction, increases JA production but restricts the accumulation of both ET and salicylic acid. JA-induced anthocyanin accumulation and chlorophyll degradation are enhanced and stomatal immunity is impaired by BIG disruption. Bacteria- and lipopolysaccaride (LPS)-induced stomatal closure is reduced in BIG gene mutants, which are hyper-susceptible to microbial pathogens with different lifestyles, but these mutants are less attractive to phytophagous insects. Our results indicate that BIG negatively and positively regulate the MYC2 and ERF1 arms of the JA signalling pathway. BIG warrants recognition as a new and distinct regulator that regulates JA responses, the synergistic interactions of JA and ET, and other hormonal interactions that reconcile the growth and defense dilemma in Arabidopsis.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/imunologia , Arabidopsis/metabolismo , Proteínas de Ligação a Calmodulina/metabolismo , Ciclopentanos/metabolismo , Oxilipinas/metabolismo , Imunidade Vegetal , Estômatos de Plantas/imunologia , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Ligação a Calmodulina/genética , Regulação para Baixo/genética , Etilenos , Regulação da Expressão Gênica de Plantas , Mutação/genética , Ácido Salicílico/metabolismo
15.
Biochim Biophys Acta Mol Cell Res ; 1866(3): 395-408, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30290240

RESUMO

Cell migration is a critical mechanism controlling tissue morphogenesis, epithelial wound healing and tumor metastasis. Migrating cells depend on orchestrated remodeling of the plasma membrane and the underlying actin cytoskeleton, which is regulated by the spectrin-adducin-based membrane skeleton. Expression of adducins is altered during tumorigenesis, however, their involvement in metastatic dissemination of tumor cells remains poorly characterized. This study investigated the roles of α-adducin (ADD1) and γ-adducin (ADD3) in regulating migration and invasion of non-small cell lung cancer (NSCLC) cells. ADD1 was mislocalized, whereas ADD3 was markedly downregulated in NSCLC cells with the invasive mesenchymal phenotype. CRISPR/Cas9-mediated knockout of ADD1 and ADD3 in epithelial-type NSCLC and normal bronchial epithelial cells promoted their Boyden chamber migration and Matrigel invasion. Furthermore, overexpression of ADD1, but not ADD3, in mesenchymal-type NSCLC cells decreased cell migration and invasion. ADD1-overexpressing NSCLC cells demonstrated increased adhesion to the extracellular matrix (ECM), accompanied by enhanced assembly of focal adhesions and hyperphosphorylation of Src and paxillin. The increased adhesiveness and decreased motility of ADD1-overexpressing cells were reversed by siRNA-mediated knockdown of Src. By contrast, the accelerated migration of ADD1 and ADD3-depleted NSCLC cells was ECM adhesion-independent and was driven by the upregulated expression of pro-motile cadherin-11. Overall, our findings reveal a novel function of adducins as negative regulators of NSCLC cell migration and invasion, which could be essential for limiting lung cancer progression and metastasis.


Assuntos
Caderinas/metabolismo , Proteínas de Ligação a Calmodulina/metabolismo , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Junções Célula-Matriz/metabolismo , Neoplasias Pulmonares/metabolismo , Caderinas/biossíntese , Caderinas/genética , Proteínas de Ligação a Calmodulina/biossíntese , Proteínas de Ligação a Calmodulina/genética , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Linhagem Celular Tumoral , Movimento Celular/fisiologia , Junções Célula-Matriz/genética , Proteínas do Citoesqueleto/genética , Proteínas do Citoesqueleto/metabolismo , Regulação para Baixo , Células Epiteliais/metabolismo , Adesões Focais/metabolismo , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Invasividade Neoplásica , RNA Interferente Pequeno/metabolismo , Transdução de Sinais
16.
Anim Biotechnol ; 30(1): 7-12, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29527980

RESUMO

The α-adducin (ADD1) is a subunit of adducin which is a cytoskeleton heterodimeric protein. Adducin participates in oocytes chromosome meiosis of mice, prompting adducin has an effect on embryonic development. Adducin gene mutation has significantly functional change. So the present study was to identify and characterize polymorphisms within the coding region of the bovine ADD1 gene among different cattle breeds. Here, 11 novel single nucleotide polymorphisms (SNPs 1-11) were identified by DNA sequencing and polymerase chain reaction-single stranded conformational polymorphism, there were one synonymous mutation in exon 1 (SNP1); four missense mutations in exons 4, 7, and 8 (SNPs 3-6); and six mutations in introns 4, 12, 13, and 14 (SNPs 2, 7-10). The statistical analyses indicated that the some SNPs are associated with the growth traits (body length, body height, chest circumference, and hucklebone width) in Chinese Jiaxian cattle population. Our results provide evidence that polymorphisms in the ADD1 gene are associated with growth traits, and may be used for marker-assisted selection in beef cattle breeding program.


Assuntos
Proteínas de Ligação a Calmodulina/genética , Bovinos/genética , Variação Genética , Animais , Tamanho Corporal/genética , Bovinos/crescimento & desenvolvimento , Éxons/genética , Fenótipo , Polimorfismo de Nucleotídeo Único/genética
17.
Am J Surg Pathol ; 43(4): 523-530, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30520818

RESUMO

NTRK fusions in malignant tumors are therapeutic targets of tyrosine kinase inhibitors. Because they occur only in a small subset of mesenchymal tumors, knowledge regarding the corresponding histology is important to effectively identify patients who could benefit from targeted therapy. In this study, using RNA sequencing, we identified novel NTRK3 fusions involving related partner genes in 2 adult bone and soft tissue tumors that met the current histologic criteria of fibrosarcoma. Case 1 involved the left radius of a 38-year-old woman, whereas in case 2, the right thigh of a 26-year-old man was affected. Histologically, both tumors consisted of the long fascicular growth of long spindle cells. The tumor in case 1 additionally showed focal myxoid changes. Tumor cells had nonpleomorphic, atypical nuclei, and lacked evidence of a specific line of differentiation. Both tumors showed widespread CD34 immunoreactivity and very limited expression of actin. RNA sequencing detected in-frame fusion transcripts of STRN (exon 3)-NTRK3 (exon 14) in case 1 and STRN3 (exon 3)-NTRK3 (exon 14) in case 2, which were confirmed by reverse transcription polymerase chain reaction and Sanger sequencing. Pan-TRK immunostaining was diffusely positive in both cases. Fluorescence in situ hybridization showed signal patterns compatible with NTRK3 rearrangements in both cases, with case 2 additionally harboring a CDKN2A homozygous deletion. This study expands the clinicopathologic and genetic spectrum of sarcomas associated with NTRK fusions, and suggests that CD34-positive fibrosarcoma of bone and soft tissue could be a good candidate for NTRK testing.


Assuntos
Autoantígenos/genética , Neoplasias Ósseas/genética , Proteínas de Ligação a Calmodulina/genética , Receptor com Domínio Discoidina 2/genética , Fibrossarcoma/genética , Proteínas de Membrana/genética , Proteínas do Tecido Nervoso/genética , Neoplasias de Tecidos Moles/genética , Adulto , Feminino , Humanos , Masculino , Fusão Oncogênica/genética , Proteínas de Fusão Oncogênica/genética
18.
J Cell Biochem ; 120(3): 4613-4619, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30552709

RESUMO

BACKGROUND: Recently, the role of α-adducin rs4961 polymorphism in hypertension (HTN) was intensively analyzed, but the results of these studies were inconsistent. Therefore, we performed this study to better assess the relationship between α-adducin rs4961 polymorphism and the likelihood of HTN. METHODS: Eligible studies were searched in PubMed, Medline, Embase, and Web of Science. Odds ratios with 95% confidence intervals were used to assess the relationship between α-adducin rs4961 polymorphism and HTN. RESULTS: A total of 33 studies with 40 432 participants were analyzed. Significant associations with the likelihood of HTN were detected for the α-adducin rs4961 polymorphism with fixed effect models (FEM) (dominant model: P = 0.003; allele model: P = 0.003), but not with random effect models (REM). Further subgroup analysis according to ethnicity of participants revealed that the α-adducin rs4961 polymorphism was significantly associated with the likelihood of HTN in Asians (7721 cases and 8299 controls) with both FEMs (dominant model: P < 0.0001; additive model: P = 0.01; allele model: P < 0.0001) and REMs (dominant model: P = 0.0005; additive model: P = 0.03; allele model: P = 0.0006). CONCLUSIONS: Our findings indicate that the α-adducin rs4961 polymorphism may serve as a genetic biomarker of HTN in Asians.


Assuntos
Grupo com Ancestrais do Continente Asiático/genética , Proteínas de Ligação a Calmodulina/genética , Hipertensão , Polimorfismo Genético , Feminino , Humanos , Hipertensão/etnologia , Hipertensão/genética , Masculino
20.
FASEB J ; 33(4): 4729-4740, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30592649

RESUMO

The adherens junctions (AJs) and tight junctions (TJs) provide critical adhesive contacts between neighboring epithelial cells and are crucial for epithelial adhesion, integrity, and barrier functions in a wide variety of tissues and organisms. The striatin protein family, which are part of the striatin interaction phosphatases and kinases complex, are multidomain scaffolding proteins that play important biologic roles. We have previously shown that striatin colocalizes with the tumor suppressor protein adenomatous polyposis coli in the TJs of epithelial cells. Here we show that striatin affects junction integrity and cell migration, probably through a mechanism that involves the adhesion molecule E-cadherin. Cells engaged in cell-cell adhesion expressed a high MW-modified form of striatin that forms stable associations with detergent-insoluble, membrane-bound cellular fractions. In addition, striatin has recently been suggested to be a target of the poly (ADP-ribose) polymerases Tankyrase 1, and we have found that striatin interacts with Tankyrase 1 and is subsequently poly-ADP-ribosylated. Taken together, our results suggest that striatin is a novel cell-cell junctional protein that functions to maintain correct cell adhesion and may have a role in establishing the relationship between AJs and TJs that is fundamental for epithelial cell-cell adhesion.-Lahav-Ariel, L., Caspi, M., Nadar-Ponniah, P. T., Zelikson, N., Hofmann, I., Hanson, K. K., Franke, W. W., Sklan, E. H., Avraham, K. B., Rosin-Arbesfeld, R. Striatin is a novel modulator of cell adhesion.


Assuntos
Proteínas de Ligação a Calmodulina/metabolismo , Adesão Celular/fisiologia , Proteínas de Membrana/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Junções Aderentes/metabolismo , Animais , Western Blotting , Células COS , Células CACO-2 , Caderinas/genética , Caderinas/metabolismo , Proteínas de Ligação a Calmodulina/genética , Adesão Celular/genética , Chlorocebus aethiops , Cães , Células Hep G2 , Humanos , Imunoprecipitação , Células MCF-7 , Células Madin Darby de Rim Canino , Proteínas de Membrana/genética , Microscopia de Fluorescência , Proteínas do Tecido Nervoso/genética , Tanquirases/metabolismo , Junções Íntimas/metabolismo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA