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1.
Physiol Mol Biol Plants ; 30(2): 337-347, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38623166

RESUMO

The vascular plant-specific type III Gγ proteins have emerged as important targets for biotechnological applications. These proteins are exemplified by Arabidopsis AGG3, rice Grain Size 3 (GS3), Dense and Erect Panicle 1 (DEP1), and GGC2 and regulate plant stature, seed size, weight and quality, nitrogen use efficiency, and multiple stress responses. These Gγ proteins are an integral component of the plant heterotrimeric G-protein complex and differ from the canonical Gγ proteins due to the presence of a long, cysteine-rich C-terminal region. Most cereal genomes encode three or more of these proteins, which have similar N-terminal Gγ domains but varying lengths of the C-terminal domain. The C-terminal domain is hypothesized to give specificity to the protein function. Intriguingly, many accessions of cultivated cereals have natural deletion of this region in one or more proteins, but the mechanistic details of protein function remain perplexing. Distinct, sometimes contrasting, effects of deletion of the C-terminal region have been reported in different crops or under varying environmental conditions. This review summarizes the known roles of type III Gγ proteins, the possible action mechanisms, and a perspective on what is needed to comprehend their full agronomic potential.

2.
Plant Cell Physiol ; 64(10): 1243-1256, 2023 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-37572092

RESUMO

The vascular plant-specific, cysteine-rich type III Gγ proteins, which are integral components of the heterotrimeric G-protein complex, play crucial roles in regulating a multitude of plant processes, including those related to crop yield and responses to abiotic stresses. The presence of multiple copies of type III Gγ proteins in most plants and a propensity of the presence of specific truncated alleles in many cultivated crops present an ambiguous picture of their roles in modulating specific responses. AGG3 is a canonical type III Gγ protein of Arabidopsis, and its overexpression in additional model crops offers the opportunity to directly evaluate the effects of protein expression levels on plant phenotypes. We have shown that AGG3 overexpression in the monocot model Setaria viridis leads to an increase in seed yield. In this study, we have investigated the response of the S. viridis plants overexpressing AGG3 to heat stress (HS), one of the most important abiotic stresses affecting crops worldwide. We show that a short span of HS at a crucial developmental time point has a significant effect on plant yield in the later stages. We also show that plants with higher levels of AGG3 are more tolerant to HS. This is attributed to an altered regulation of stress-responsive genes and improved modulation of the photosynthetic efficiency during the stress. Overall, our results confirm that AGG3 plays a crucial role in regulating plant responses to unfavorable environmental conditions and may contribute positively to avoiding crop yield losses.


Assuntos
Arabidopsis , Setaria (Planta) , Arabidopsis/genética , Arabidopsis/metabolismo , Resposta ao Choque Térmico/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Setaria (Planta)/genética , Setaria (Planta)/metabolismo , Estresse Fisiológico/genética
3.
Int J Mol Sci ; 24(10)2023 May 12.
Artigo em Inglês | MEDLINE | ID: mdl-37240005

RESUMO

Prions are transmissible self-perpetuating protein isoforms associated with diseases and heritable traits. Yeast prions and non-transmissible protein aggregates (mnemons) are frequently based on cross-ß ordered fibrous aggregates (amyloids). The formation and propagation of yeast prions are controlled by chaperone machinery. Ribosome-associated chaperone Hsp70-Ssb is known (and confirmed here) to modulate formation and propagation of the prion form of the Sup35 protein [PSI+]. Our new data show that both formation and mitotic transmission of the stress-inducible prion form of the Lsb2 protein ([LSB+]) are also significantly increased in the absence of Ssb. Notably, heat stress leads to a massive accumulation of [LSB+] cells in the absence of Ssb, implicating Ssb as a major downregulator of the [LSB+]-dependent memory of stress. Moreover, the aggregated form of Gγ subunit Ste18, [STE+], behaving as a non-heritable mnemon in the wild-type strain, is generated more efficiently and becomes heritable in the absence of Ssb. Lack of Ssb also facilitates mitotic transmission, while lack of the Ssb cochaperone Hsp40-Zuo1 facilitates both spontaneous formation and mitotic transmission of the Ure2 prion, [URE3]. These results demonstrate that Ssb is a general modulator of cytosolic amyloid aggregation, whose effect is not restricted only to [PSI+].


Assuntos
Subunidades gama da Proteína de Ligação ao GTP , Príons , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Chaperonas Moleculares/metabolismo , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP70/metabolismo , Príons/metabolismo , Glutationa Peroxidase/metabolismo , Subunidades gama da Proteína de Ligação ao GTP/metabolismo , Fatores de Terminação de Peptídeos/metabolismo
4.
Hemoglobin ; 46(6): 317-324, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36815306

RESUMO

ß-thalassemia is a serious public health problem in Sabah due to its high prevalence. This study aimed to investigate the effects of different types of ß-globin gene mutations, coinheritance with α-globin gene mutations, XmnI-Gγ, and rs368698783 polymorphisms on the ß-thalassemia phenotypes in Sabahan patients. A total of 111 patients were included in this study. The sociodemographic profile of the patients was collected using a semi-structured questionnaire, while clinical data were obtained from their medical records. Gap-PCR, ARMS-PCR, RFLP-PCR, and multiplex PCR were performed to detect ß- and α-globin gene mutations, as well as XmnI-Gγ and rs368698783 polymorphisms. Our data show that the high prevalence of ß-thalassemia in Sabah is not due to consanguineous marriages (5.4%). A total of six different ß-globin gene mutations were detected, with Filipino ß°-deletion being the most dominant (87.4%). There were 77.5% homozygous ß-thalassemia patients, 16.2% compound heterozygous ß-thalassemia patients, and 6.3% ß-thalassemia/Hb E patients. Further evaluation on compound heterozygous ß-thalassemia and ß-thalassemia/Hb E patients found no concomitant α-globin gene mutations and the rs368698783 polymorphism. Furthermore, the XmnI-Gγ (-/+) genotype did not demonstrate a strong impact on the disease phenotype, as only two of five patients in the compound heterozygous ß-thalassemia group and two of three patients in the ß-thalassemia/Hb E group had a moderate phenotype. Our findings indicate that the severity of the ß-thalassemia phenotypes is closely related to the type of ß-globin gene mutations but not to the XmnI-Gγ and rs368698783 polymorphisms.


Assuntos
Talassemia , Talassemia beta , Humanos , Talassemia beta/genética , Malásia , Mutação , Genótipo , Fenótipo , alfa-Globinas/genética , Reação em Cadeia da Polimerase Multiplex , Globinas beta/genética
5.
Plant Mol Biol ; 106(6): 505-520, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34176052

RESUMO

KEY MESSAGE: Gene expression analysis coupled with in-planta studies showed that specific Gßγ combination regulates plant growth and defence traits in the allotetraploid Brassica juncea. Plant heterotrimeric G-proteins regulate a wide range of responses despite their limited repertoire of core components. The roles and functional interactions between different G-protein subunits are quite perplexing, which get further complicated with polyploidy. Here, we show that the allotetraploid Brassica juncea comprises multiple homologs of G-protein genes, encoding six BjuGß and ten highly divergent BjuGγ subunit proteins, later being classified into type-A1, type-A2 and type-C Gγ proteins. The encoded BjuGß and BjuGγ proteins shared close evolutionary relationship and have retained distinct spatio-temporal expression patterns during plant developmental stages and in response to the necrotrophic pathogen, Sclerotinia sclerotiorum. RNAi based suppression of BjuGß and BjuGγ genes suggested functional overlap and selectivity of BjuGßs with three distinct BjuGγ type subunits, to regulate plant height (BjuGßγA2 and BjuGßγC), seed weight (BjuGßGγA1 and BjuGßGγC), silique size (BjuGßGγC) and pathogen response (BjuGßGγA1 and BjuGßGγC). Further, the triplicated BjuGß genes, formed due to Brassica specific whole-genome-triplication event, showed differential involvement during pathogen response, wherein overexpression of BjuGß2 displayed higher resistance to Sclerotinia infection. Taken together, our study demonstrates that multiple BjuGß and BjuGγ proteins have retained distinct spatio-temporal expression and functional selectivity to regulate specific plant growth and defence traits in the oilseed B. juncea.


Assuntos
Subunidades beta da Proteína de Ligação ao GTP/genética , Subunidades gama da Proteína de Ligação ao GTP/genética , Mostardeira/genética , Doenças das Plantas/genética , Proteínas de Plantas/genética , Poliploidia , Ascomicetos/fisiologia , Resistência à Doença/genética , Subunidades beta da Proteína de Ligação ao GTP/classificação , Subunidades beta da Proteína de Ligação ao GTP/metabolismo , Subunidades gama da Proteína de Ligação ao GTP/classificação , Subunidades gama da Proteína de Ligação ao GTP/metabolismo , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica de Plantas , Interações Hospedeiro-Patógeno , Modelos Genéticos , Mostardeira/crescimento & desenvolvimento , Mostardeira/microbiologia , Filogenia , Doenças das Plantas/microbiologia , Proteínas de Plantas/classificação , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Ligação Proteica , Interferência de RNA
6.
BMC Plant Biol ; 21(1): 471, 2021 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-34654373

RESUMO

BACKGROUND: Heterotrimeric G-proteins, composed of Gα, Gß and Gγ subunits, are important signal transmitters, mediating the cellular response to multiple stimuli in animals and plants. The Gγ subunit is an essential component of the G-protein, providing appropriate functional specificity to the heterotrimer complex and has been well studied in many species. However, the evolutionary history, expression pattern and functional characteristics of Gγ subunits has not been explored in the Rosaceae, representing many important fruit crops. RESULTS: In this study, 35 Gγ subunit genes were identified from the eight species belonging to the Rosaceae family. Based on the structural gene characteristics, conserved protein motifs and phylogenetic analysis of the Gγ subunit genes, the genes were classified into three clades. Purifying selection was shown to play an important role in the evolution of Gγ subunit genes, while a recent whole-genome duplication event was the principal force determining the expansion of the Gγ subunit gene family in the subfamily Maloideae. Gγ subunit genes exhibited diverse spatiotemporal expression patterns in Chinese white pear, including fruit, root, ovary and bud, and under abiotic stress conditions, the relative expression of Gγ subunit genes were up-regulated or down-regulated. In addition, seven of the Gγ subunit proteins in pear were located on the plasma membrane, in the cytoplasm or nucleus. CONCLUSION: Overall, this study of the Gγ subunit gene family in eight Rosaceae species provided useful information to better understand the evolution and expression of these genes and facilitated further exploration of their functions in these important crop plants.


Assuntos
Subunidades gama da Proteína de Ligação ao GTP/genética , Genoma de Planta/genética , Família Multigênica/genética , Pyrus/genética , Rosaceae/genética , Motivos de Aminoácidos , Evolução Molecular , Perfilação da Expressão Gênica , Filogenia , Proteínas de Plantas/genética , Transdução de Sinais , Estresse Fisiológico
7.
J Korean Med Sci ; 35(34): e281, 2020 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-32864904

RESUMO

On October 4, 2018, an outbreak of gastroenteritis associated with sapovirus occurred among elementary school students in Gyeonggi-do, Korea. Epidemiologic studies were conducted in a retrospective cohort approach. Using self-administered questionnaires, we collected information on symptoms and food items consumed. Of the 999 subjects, 17 developed patients that met the case definition. The main symptom was vomiting (100%), and the symptomatic age was 6-12 years. Positive samples were identified by conventional reverse transcription polymerase chain reaction for sequencing. They were classified into genotype GI.3 by phylogenetic analysis. This is the first report of an outbreak associated with sapovirus GI.3 in Korea.


Assuntos
Infecções por Caliciviridae/diagnóstico , Gastroenterite/diagnóstico , Sapovirus/isolamento & purificação , Infecções por Caliciviridae/epidemiologia , Infecções por Caliciviridae/virologia , Criança , Surtos de Doenças , Feminino , Gastroenterite/epidemiologia , Gastroenterite/virologia , Genótipo , Humanos , Masculino , Filogenia , RNA Viral/análise , República da Coreia/epidemiologia , Estudos Retrospectivos , Sapovirus/classificação , Sapovirus/genética , Instituições Acadêmicas
8.
J Cell Sci ; 129(16): 3153-66, 2016 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-27390173

RESUMO

Extracellular ligands control biological phenomena. Cells distinguish physiological stimuli from weak noise stimuli by establishing a ligand-concentration threshold. Hormonal control of the meiotic G2/M transition in oocytes is essential for reproduction. However, the mechanism for threshold establishment is unclear. In starfish oocytes, maturation-inducing hormones activate the PI3K-Akt pathway through the Gßγ complex of heterotrimeric G-proteins. Akt directly phosphorylates both Cdc25 phosphatase and Myt1 kinase, resulting in activation of cyclin-B-Cdk1, which then induces meiotic G2/M transition. Here, we show that cyclin-B-Cdk1 is partially activated after subthreshold hormonal stimuli, but this triggers negative feedback, resulting in dephosphorylation of Akt sites on Cdc25 and Myt1, thereby canceling the signal. We also identified phosphatase activity towards Akt substrates that exists independent of stimuli. In contrast to these negative regulatory activities, an atypical Gßγ-dependent pathway enhances PI3K-Akt-dependent phosphorylation. Based on these findings, we propose a model for threshold establishment in which hormonal dose-dependent competition between these new pathways establishes a threshold; the atypical Gßγ-pathway becomes predominant over Cdk-dependent negative feedback when the stimulus exceeds this threshold. Our findings provide a regulatory connection between cell cycle and signal transduction machineries.


Assuntos
Proteína Quinase CDC2/metabolismo , Ciclina B/metabolismo , Fase G2 , Meiose , Mitose , Estrelas-do-Mar/citologia , Estrelas-do-Mar/metabolismo , Adenina/análogos & derivados , Adenina/farmacologia , Animais , Ativação Enzimática/efeitos dos fármacos , Retroalimentação Fisiológica/efeitos dos fármacos , Fase G2/efeitos dos fármacos , Subunidades beta da Proteína de Ligação ao GTP/metabolismo , Humanos , Meiose/efeitos dos fármacos , Mitose/efeitos dos fármacos , Modelos Biológicos , Proteínas Mutantes/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação/efeitos dos fármacos , Fosfosserina/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Estrelas-do-Mar/efeitos dos fármacos , Especificidade por Substrato/efeitos dos fármacos , Fosfatases cdc25/metabolismo
9.
Cell Mol Biol (Noisy-le-grand) ; 64(1): 32-39, 2018 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-29412791

RESUMO

In the present study, sixty-two samples that have 1.5% and upper level of fetal hemoglobin (HbF), were examined to investigate the relationship between HbF level and non-deletional mutations in both Gγ (G gamma) globin (HBG2) and Aγ (A gamma) globin (HBG1) genes. Four variations were observed in the promotor of Gγ gene, which are -158C/T, -309A/G, -369C/G, and -567T/G. Also, four variations were observed in the 5'-UTR (untranslated regions) and promotor of Aγ gene, which are +25G/A, -369G/C, -499T/A, and -588G/A. One -222/-225 AGCA del homozygous and six variations as heterozygous in A gamma globin gene promotor region were also observed. The results of the current study suggested that there was a significant relationship between high HbF levels and two variations (-309A/T and -369C/G) in Gγ gene promotor. Additionally, a significant relationship between two variations (+25G/A and -499T/A) in Aγ gene promotor was also observed. Furthermore, the persons who carry these variations with high levels of HbF indicated that there might be a haplotype effect between these variations.


Assuntos
Hemoglobina Fetal/metabolismo , Variação Genética , Regiões Promotoras Genéticas , gama-Globinas/genética , Humanos , Padrões de Herança
10.
Plant J ; 81(3): 388-98, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25430066

RESUMO

Heterotrimeric G proteins are crucial for the perception of external signals and subsequent signal transduction in animal and plant cells. In both model systems, the complex comprises one Gα, one Gß, and one Gγ subunit. However, in addition to the canonical Gγ subunits (class A), plants also possess two unusual, plant-specific classes of Gγ subunits (classes B and C) that have not yet been found in animals. These include Gγ subunits lacking the C-terminal CaaX motif (class B), which is important for membrane anchoring of the protein; the presence of such subunits gives rise to a flexible sub-population of Gß/γ heterodimers that are not necessarily restricted to the plasma membrane. Plants also contain class C Gγ subunits, which are twice the size of canonical Gγ subunits, with a predicted transmembrane domain and a large cysteine-rich extracellular C-terminus. However, neither the presence of the transmembrane domain nor the membrane topology have been unequivocally demonstrated. Here, we provide compelling evidence that AGG3, a class C Gγ subunit of Arabidopsis, contains a functional transmembrane domain, which is sufficient but not essential for plasma membrane localization, and that the cysteine-rich C-terminus is extracellular.


Assuntos
Proteínas de Arabidopsis/química , Arabidopsis/metabolismo , Subunidades gama da Proteína de Ligação ao GTP/química , Arabidopsis/genética , Proteínas de Arabidopsis/análise , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/fisiologia , Membrana Celular/metabolismo , Subunidades alfa de Proteínas de Ligação ao GTP/genética , Subunidades alfa de Proteínas de Ligação ao GTP/metabolismo , Subunidades alfa de Proteínas de Ligação ao GTP/fisiologia , Subunidades beta da Proteína de Ligação ao GTP/genética , Subunidades beta da Proteína de Ligação ao GTP/metabolismo , Subunidades beta da Proteína de Ligação ao GTP/fisiologia , Subunidades gama da Proteína de Ligação ao GTP/análise , Subunidades gama da Proteína de Ligação ao GTP/genética , Modelos Moleculares , Filogenia , Estrutura Terciária de Proteína , Análise de Sequência de Proteína
11.
Hemoglobin ; 40(1): 61-3, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26573261

RESUMO

Here we describe Hb F-Avellino [(G)γ41(C7)Phe → Leu; HBG2: c.124 T > C], a new hemoglobin (Hb) variant observed in a healthy newborn. The proband's hemolysate was found to be mildly unstable by the isopropanol test. The occurrence of the variant was assessed by both chromatographic and electrophoretic methods. DNA sequencing analysis of the (G)γ gene showed a T to C transition at codon 41 (TTC > CTC) corresponding to the Phe → Leu substitution. Normal functional properties have been hypothesized.


Assuntos
Hemoglobina Fetal/genética , Mutação Puntual , gama-Globinas/genética , Substituição de Aminoácidos , Códon , Variação Genética , Humanos , Recém-Nascido
12.
Curr Genet ; 61(4): 641-51, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25944571

RESUMO

Heterotrimeric G-proteins play key roles in the transduction of extracellular signals to various downstream effectors in eukaryotes. In our previous study, a T-DNA insertional mutant A1-412, in which the promoter of a putative Gγ subunit gene MGG1 was disrupted, was impaired in asexual/sexual sporulation, appressorium formation, and pathogenicity in Magnaporthe oryzae. Here the roles of MGG1 in regulating fungal development and plant infection were further investigated and verified using a gene deletion strategy. Targeted gene deletion mutants of MGG1 exhibited similar phenotypes to those of A1-412. The Δmgg1 mutants were unable to differentiate appressorium on hydrophobic surfaces and nonpathogenic to susceptible hosts. The defects of the Δmgg1 mutants in appressorium formation were partially restored by adding exogenous cAMP or IBMX (a phosphodiesterase inhibitor), although the induced appressoria were still nonfunctional. Expressing Mgg1-GFP fusion protein in an Δmgg1 mutant could complement all phenotypes of the mutant, and bright GFP fluorescence was observed at the periphery of fungal cells, indicating that Mgg1 mainly localizes to plasma membrane. Quantitative RT-PCR analysis revealed that deletion of MGG1 resulted in a significant reduction in mRNA levels of the genes encoding Gα (MagA, MagB, and MagC), Gß (Mgb1), and adenylate cyclase (Mac1). Moreover, intracellular cAMP accumulation was significantly reduced in Δmgg1 mutants compared to that in the wild-type strain. Taken together, our results suggested that Gγ subunit Mgg1 might act upstream of cAMP signaling pathway and play critical roles in regulation of conidiation, appressorium formation, mating, and plant infection in M. oryzae.


Assuntos
Proteínas Fúngicas/genética , Subunidades gama da Proteína de Ligação ao GTP/genética , Regulação Fúngica da Expressão Gênica , Magnaporthe/genética , Magnaporthe/patogenicidade , 1-Metil-3-Isobutilxantina/farmacologia , AMP Cíclico/metabolismo , AMP Cíclico/farmacologia , DNA Bacteriano/genética , DNA Bacteriano/metabolismo , Proteínas Fúngicas/metabolismo , Subunidades alfa de Proteínas de Ligação ao GTP/genética , Subunidades alfa de Proteínas de Ligação ao GTP/metabolismo , Subunidades gama da Proteína de Ligação ao GTP/deficiência , Deleção de Genes , Genes Fúngicos Tipo Acasalamento , Teste de Complementação Genética , Hifas/genética , Hifas/metabolismo , Hifas/patogenicidade , Magnaporthe/metabolismo , Oryza/microbiologia , Fenótipo , Inibidores de Fosfodiesterase/farmacologia , Doenças das Plantas/microbiologia , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Transdução de Sinais , Esporos Fúngicos/genética , Esporos Fúngicos/metabolismo , Esporos Fúngicos/patogenicidade , Virulência
13.
Stress Biol ; 3(1): 9, 2023 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-37676334

RESUMO

This brief article highlights the results of Zhang et al. (Science 379, eade8416, 2023), who recently found that the Gγ subunit AT1/GS3 contributes to alkaline tolerance in several main monocots crops, and revealed the molecular mechanism of AT1/GS3-mediated response to alkaline stress in plants, which involves regulating H2O2 levels by inhibiting the phosphorylation of aquaporin PIP2s.

14.
Genes (Basel) ; 11(8)2020 08 09.
Artigo em Inglês | MEDLINE | ID: mdl-32784857

RESUMO

Recombination is one of the major sources of genetic variation in viruses. RNA viruses, such as rabbit hemorrhagic disease virus (RHDV), are among the viruses with the highest recombination rates. Several recombination events have been described for RHDV, mostly as a consequence of their genomic architecture. Here, we undertook phylogenetic and recombination analyses of French and Swedish RHDV strains from 1994 to 2016 and uncovered a new intergenotypic recombination event. This event occurred in the late 1990s/early 2000s and involved nonpathogenic GI.3 strains as donors for the nonstructural part of the genome of these recombinants, while pathogenic GI.1d strains contributed to the structural part. These GI.3P-GI.1d recombinant strains did not entirely replace GI.1d (nonrecombinant) strains, but became the dominant strains in France and Sweden, likely due to a fitness advantage associated with this genomic architecture. GI.3P-GI.1d (P stands for polymerase) strains persisted until 2013 and 2016 in Sweden and France, respectively, and cocirculated with the new genotype GI.2 in France. Since strains from the first GI.2 outbreaks were GI.3P-GI.2, we hypothesize that GI.3P-GI.1d could be the parental strain. Our results confirm the outstanding recombination ability of RHDV and its importance in the evolution of lagoviruses, which was only revealed by studying complete genomic sequences.


Assuntos
Doenças dos Animais/epidemiologia , Doenças dos Animais/virologia , Infecções por Caliciviridae/veterinária , Vírus da Doença Hemorrágica de Coelhos/classificação , Vírus da Doença Hemorrágica de Coelhos/genética , Recombinação Genética , Animais , Animais Selvagens , Evolução Molecular , França/epidemiologia , Genoma Viral , Genótipo , História do Século XX , Filogenia , RNA Viral , Estudos Retrospectivos , Suécia/epidemiologia
15.
Infect Genet Evol ; 85: 104554, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32927119

RESUMO

Noroviruses (NoVs) are a major cause of acute non-bacterial gastroenteritis worldwide. In this study, we report the isolation, near-complete genome sequencing, and expression and biological characterization of the major capsid protein (VP1) of a GI.3 NoV isolated from a child presenting acute gastroenteritis. The genome of the GI.3 NoV is 7746 bp in length, not including the poly-adenylation tail. Phylogenetic analysis based on the complete VP1 nucleotide sequences indicates that GI.3 NoVs could be divided into four clusters, with 4.6%, 5.3%, 6.6%, 1.9% intracluster variations in nucleotide and 4.8%, 3.8%, 6.1%, 1.7% intracluster variations in amino acid sequences, respectively. A Bayesian evolutionary analysis showed that GI.3 NoVs evolved at 2.44 × 10-3, 2.78 × 10-3, and 3.04 × 10-3 nucleotide substitutions/site/year using a strict clock model, an uncorrelated log-normal model (UCLN), and an uncorrelated exponential derivation model (UCED), respectively. VP1 protein expression using a recombinant baculovirus expression system leads to the successful assembly of virus-like particles (VLPs). In vitro VLP-Histo-blood group antigen (HBGA) binding assay indicates that GI.3 NoV VLPs strongly bind to blood type A salivary HBGAs, moderately bind to blood type O salivary HBGAs, and weakly bind or do not bind to blood type B and AB salivary HBGAs. In vitro VLP-HBGA binding blockade assay indicated that the binding of GI.3 NoV VLPs to blood type A salivary HBGAs could only be blocked by anti-GI.3 NoV VLPs serum but not non-GI.3 NoV genotype-specific hyperimmune sera (GI.2, GI.7, GII.4, GII.6, GII.7, and GII.17). The detailed characterization of GI.3 NoV in this study provides evidence that GI.3 NoV undergoes rapid evolution and exhibits no cross-blocking effects, suggesting that GI.3 NoV may potentially be utilized in the development of multivalent NoV vaccines.


Assuntos
Infecções por Caliciviridae/virologia , Gastroenterite/virologia , Norovirus/classificação , Norovirus/genética , Sequência de Aminoácidos , Sequência de Bases , Infecções por Caliciviridae/imunologia , Infecções por Caliciviridae/prevenção & controle , Gastroenterite/imunologia , Gastroenterite/prevenção & controle , Genoma Viral , Genômica/métodos , Interações Hospedeiro-Patógeno/imunologia , Humanos , Norovirus/imunologia , Filogenia , Ligação Proteica , Vacinas de Partículas Semelhantes a Vírus/imunologia , Proteínas Virais/imunologia , Proteínas Virais/metabolismo , Vacinas Virais/imunologia
16.
Artigo em Inglês | MEDLINE | ID: mdl-32411617

RESUMO

Brucella abortus is a facultative intracellular pathogen that causes a zoonosis called brucellosis. This disease leads to abortion and infertility in cattle, and diverse complications in humans. B. abortus is a successful intracellular bacterium that has developed the ability to evade the host's immune system and it replicates in professional and non-professional phagocytic cells, persisting in the different tissues, and organs of its hosts. It has been described that Brucella expresses a polar flagellum under certain conditions, but its function is still unknown. In this study we evaluated the role of the FlgJ, a protein, presumably a peptidoglycan hydrolase involved in flagellum formation and in the virulence of B. abortus strain 2308. B. abortus 2308 ΔflgJ mutant and complemented strains were constructed to study the function of the FlgJ protein in the context of the virulence of this pathogen in in vitro and in vivo assays. The results showed that the elimination of the flgJ gene delays the growth rate of B. abortus in culture, reduces its intracellular survival capacity in professional and non-professional phagocytic cells, rendering it unable to escape from the endocytic route and not reaching the endoplasmic reticulum. It also negatively affects their persistence in BALB/c mice. Functionally, the B. abortus 2308 flgJ gene restored motility to an E. coli flgJ mutant gene. Furthermore, it was discovered that the production of FlgJ protein is associated with the bacterial adherence by B. abortus. Therefore, although the specific function of the polar flagellum for Brucella is unknown, the data indicates that the flagellar flgJ gene and its product are required for full virulence of B. abortus 2308, since its deletion significantly reduces the fitness of this pathogen in vitro and in vivo.


Assuntos
Brucella abortus , Brucelose , Animais , Brucella abortus/genética , Bovinos , Escherichia coli , Camundongos , Camundongos Endogâmicos BALB C , Virulência
17.
Mediterr J Hematol Infect Dis ; 11(1): e2019038, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31308914

RESUMO

BACKGROUND: The finding of many Thai Hb E-ß0-thalassemia patients with non-transfusion dependent thalassemia (NTDT) phenotype without co-inheritance of α-thalassemia has prompted us to investigate the existence of other genetic modifying factors. METHODS: Study was done on 122 adult Thai patients with NTDT Hb E-ß-thalassemia patients without co-inheritance of α-thalassemia. Multiple single-nucleotide polymorphisms (SNPs) associated with γ-globin gene expression including the Gγ-XmnI of HBG2 gene, rs2297339, rs4895441, and rs9399137 of the HBS1L-MYB gene, rs4671393 in the BCL11A gene, and G176AfsX179, T334R, R238H and -154 (C-T) in the KLF1 gene were investigated using PCR and related techniques. RESULTS: Heterozygous and homozygous for Gγ-XmnI of HBG2 gene were detected at 70.5% and 7.4%, respectively. Further DNA analysis identified the rs2297339 (C-T), rs4895441 (A-G), and rs9399137 (T-C) of HBS1L-MYB gene in 86.9%, 25.4%, and 23.0%, respectively. The rs4671393 (G-A) of the BCL11A gene was found at 31.2%. For the KLF1 gene, only T334R was detected at 9.0%. CONCLUSIONS: It was found that these SNPs, when analyzed in combination, could explain the mild phenotypic expression of all cases. These results underline the importance of these informative SNPs on phenotypic expression of Hb E-ß-thalassemia patients.

18.
Plant Sci ; 270: 292-300, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29576082

RESUMO

Heterotrimeric G-proteins are key signaling components involved during the regulation of a multitude of growth and developmental pathways in all eukaryotes. Although the core proteins (Gα, Gß, Gγ subunits) and their basic biochemistries are conserved between plants and non-plant systems, seemingly different inherent properties of specific components, altered wirings of G-protein network architectures, and the presence of novel receptors and effector proteins make plant G-protein signaling mechanisms somewhat distinct from the well-established animal paradigm. G-protein research in plants is getting a lot of attention recently due to the emerging roles of these proteins in controlling many agronomically important traits. New findings on both canonical and novel G-protein components and their conserved and unique signaling mechanisms are expected to improve our understanding of this important module in affecting critical plant growth and development pathways and eventually their utilization to produce plants for the future needs. In this review, we briefly summarize what is currently known in plant G-protein research, describe new findings and how they are changing our perceptions of the field, and discuss important issues that still need to be addressed.


Assuntos
Proteínas Heterotriméricas de Ligação ao GTP/metabolismo , Desenvolvimento Vegetal , Fenômenos Fisiológicos Vegetais , Plantas , Transdução de Sinais , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/fisiologia , Proteínas Heterotriméricas de Ligação ao GTP/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
19.
Front Plant Sci ; 9: 109, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29479357

RESUMO

Heterotrimeric G-proteins are key regulators of a multitude of growth and development pathways in eukaryotes. Along with the conserved G-protein components found in all organisms, plants have certain novel variants with unique architecture, which may be involved in the regulation of plant-specific traits. The higher plant-specific type III (or Class C) Gγ protein, which possesses a large C terminal extension, represented by AGG3 in Arabidopsis, is one such variant of canonical Gγ proteins. The type III Gγ proteins are involved in regulation of many agronomically important traits in plants, including seed yield, organ size regulation, abscisic acid (ABA)-dependent signaling and stress responses, and nitrogen use efficiency. However, the extant data, especially in the monocots, present a relatively complex and sometimes contradictory picture of the regulatory role of these proteins. It remains unclear if the positive traits observed in certain naturally occurring populations are due to the presence of specific allelic variants of the proteins or due to the altered expression of the gene itself. To address these possibilities, we have overexpressed the Arabidopsis AGG3 gene in the model monocot Setaria viridis and systematically evaluated its role in conferring agriculturally relevant phenotypes. Our data show that AtAGG3 is indeed functional in Setaria and suggest that a subset of the traits affected by the type III Gγ proteins are indeed positively correlated with the gene expression level, while others might have more complex, allele specific regulation.

20.
Biol Bull ; 233(1): 58-69, 2017 08.
Artigo em Inglês | MEDLINE | ID: mdl-29182505

RESUMO

Visual pigments, each composed of an opsin protein covalently bound to a chromophore molecule, confer light sensitivity for vision. The eyes of some species of stomatopod crustaceans, or mantis shrimp, can express dozens of different opsin genes. The opsin diversity, along with spectral filters and unique tripartite eye structure, bestow upon stomatopods unusually complex visual systems. Although opsins are found in tissues outside typical image-forming eyes in other animals, extraocular opsin expression in stomatopods, animals well known for their diversity of opsins, was unknown. Caudal photoreception in the central nervous system of decapod crustaceans, a group closely related to stomatopod crustaceans, is thought to be opsin based. However, electrophysiological data suggest that stomatopods do not have caudal photoreceptors. In this study, we identified mRNAs that could encode four different opsins and several components of a potential Gq-mediated phototransduction pathway in the central nervous system of the Caribbean mantis shrimp Neogonodactylus oerstedii. The four opsins are abundantly expressed in the cerebral ganglion, or brain, with little or no expression in the remainder of the ventral nerve cord. Our data suggest that there are previously undiscovered cerebral photoreceptors in stomatopods.


Assuntos
Decápodes/genética , Expressão Gênica , Opsinas/genética , Animais , Encéfalo/fisiologia , Região do Caribe , Decápodes/fisiologia , Transcriptoma
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