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1.
Plant Sci ; 306: 110874, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33775370

RESUMO

EARLY BUD-BREAK 1 (EBB1) can promote bud break, and this function is likely conserved in woody plants. To get a more comprehensive understand of its function, peach (Prunus persica var. nectarina cultivar Zhongyou 4) PpEBB1 was overexpressed in Arabidopsis; the resultant phenotypes, including curved leaves, abnormal development of floral organs and low seed set, were similar to those of DORNRÖSCHEN-LIKE (DRNL) overexpression, indicating that PpEBB1 was a putative ortholog of AtDRNL. PpEBB1 bound to the GCC box-like element in the STYLISH1/SHI RELATED SEQUENCE5 (STY1/SRS5) promoter of peach, which has been proposed to occur in Arabidopsis as well. A GCC box-like element was also found in the YUCCA1 (YUC1) promoter, and PpEBB1 could bind to this element and activate the expression of YUC1. In addition to the elevated auxin content in the PpEBB1-oe plants as observed in our previous study, these results suggest that PpEBB1 can regulate auxin biosynthesis by directly activating related genes. Besides, we screened a zinc finger RING-finger protein, MYB30-INTERACTING E3 LIGASE 1 (PpMIEL1), showing interaction with PpEBB1, suggesting that the stability of PpEBB1 might be influenced by PpMIEL1 through ubiquitination.


Assuntos
Flores/crescimento & desenvolvimento , Flores/genética , Ácidos Indolacéticos/metabolismo , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/genética , Prunus persica/crescimento & desenvolvimento , Prunus persica/genética , Proteínas de Transporte/genética , Proteínas de Transporte/fisiologia , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Regiões Promotoras Genéticas , Fatores de Transcrição
2.
Crit Care ; 25(1): 53, 2021 02 08.
Artigo em Inglês | MEDLINE | ID: mdl-33557908

RESUMO

The current pandemic of COVID-19 caused thousands of deaths and healthcare professionals struggle to properly manage infected patients. This review summarizes information about SARS-CoV-2 receptor binding dynamics and intricacies, lung autopsy findings, immune response patterns, evidence-based explanations for the immune response, and COVID-19-associated hypercoagulability.


Assuntos
/fisiopatologia , Proteínas de Transporte/fisiologia , Pneumopatias/fisiopatologia , Pneumonia Viral/fisiopatologia , /patogenicidade , /imunologia , Proteínas de Transporte/imunologia , Humanos , Pneumopatias/imunologia , Pandemias , Pneumonia Viral/imunologia , /imunologia
3.
Dev Biol ; 469: 37-45, 2021 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-33022230

RESUMO

How organisms control organ size is not fully understood. We found that Syd/JIP3 is required for proper wing size in Drosophila. JIP3 mutations are associated with organ size defects in mammals. The underlying mechanisms are not well understood. We discovered that Syd/JIP3 inhibition results in a downregulation of the inhibitor of apoptosis protein 1 (Diap1) in the Drosophila wing. Correspondingly, Syd/JIP3 deficient tissues exhibit ectopic cell death and yield smaller wings. Syd/JIP3 inhibition generated similar effects in mammalian cells, indicating a conserved mechanism. We found that Yorkie/YAP stimulates Syd/JIP3 in Drosophila and mammalian cells. Notably, Syd/JIP3 is required for the full effect of Yorkie-mediated tissue growth. Thus Syd/JIP3 regulation of Diap1 functions downstream of Yorkie/YAP to control growth. This study provides mechanistic insights into the recent and perplexing link between JIP3 mutations and organ size defects in mammals, including in humans where de novo JIP3 variants are associated with microcephaly.


Assuntos
Proteínas de Transporte/fisiologia , Proteínas de Drosophila/metabolismo , Proteínas de Drosophila/fisiologia , Proteínas Inibidoras de Apoptose/metabolismo , Proteínas de Membrana/fisiologia , Asas de Animais/crescimento & desenvolvimento , Animais , Proteínas de Transporte/genética , Drosophila/anatomia & histologia , Drosophila/crescimento & desenvolvimento , Drosophila/metabolismo , Proteínas de Drosophila/genética , Feminino , Técnicas de Silenciamento de Genes , Células HEK293 , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Proteínas de Membrana/genética , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Proteínas Nucleares/metabolismo , Tamanho do Órgão , Proteínas Serina-Treonina Quinases/metabolismo , Transativadores/metabolismo , Asas de Animais/anatomia & histologia
4.
Life Sci ; 266: 118938, 2021 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-33347878

RESUMO

Oxidative stress is a promoting factor in the pathologic process of glucocorticoid - induced osteoporosis (GIO), while the mechanism is still unclear. Thioredoxin-interacting protein (TXNIP) is a vital protein responsible for regulation of cellular reactive oxygen species (ROS) generation elicited by mitochondrial oxidative stress, and which may activate oxidative phosphorylation under the pathogenic status. In this research, the results showed that signaling pathway associated with the mitochondrial oxidative phosphorylation (MOP) down-regulated under conditions of TXNIP siRNA in MG63 cells. Furthermore, the evidence revealed that the expression level of TXNIP in serum and bone was elevated in a rat of GIO. Moreover, the differential proteins (Ndufs3, SDHD, Cyt B, COX IV, and ATP B) related to MOP pathway were identified to down-regulate in the proteomics of bone tissues by using isobaric Tags for Relative and Absolute Quantification (iTRAQ) method in TXNIP knockout mice treated with glucocorticoid, and the proteins were also verified by simple western blot. Taken together, the present findings highlights that TXNIP involves in triggering the process of bone loss via up-regulation of the MOP pathway, resulting to GIO, while TXNIP knockout can prevent the pathogenesis of GIO to some extent.


Assuntos
Reabsorção Óssea/etiologia , Proteínas de Transporte/fisiologia , Proteínas de Ciclo Celular/metabolismo , Glucocorticoides/toxicidade , Mitocôndrias/patologia , Osteoporose/patologia , Fosforilação Oxidativa , Tiorredoxinas/fisiologia , Animais , Reabsorção Óssea/metabolismo , Reabsorção Óssea/patologia , Proteínas de Ciclo Celular/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Mitocôndrias/metabolismo , Osteoporose/induzido quimicamente , Osteoporose/metabolismo , Estresse Oxidativo , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais
6.
Nat Cell Biol ; 22(6): 663-673, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32393887

RESUMO

The linear ubiquitin chain assembly complex (LUBAC), which consists of HOIP, SHARPIN and HOIL-1L, promotes NF-κB activation and protects against cell death by generating linear ubiquitin chains. LUBAC contains two RING-IBR-RING (RBR) ubiquitin ligases (E3), and the HOIP RBR is responsible for catalysing linear ubiquitination. We found that HOIL-1L RBR plays a crucial role in regulating LUBAC. HOIL-1L RBR conjugates monoubiquitin onto all LUBAC subunits, followed by HOIP-mediated conjugation of linear chains onto monoubiquitin, and these linear chains attenuate the functions of LUBAC. The introduction of E3-defective HOIL-1L mutants into cells augmented linear ubiquitination, which protected the cells against Salmonella infection and cured dermatitis caused by reduced LUBAC levels due to SHARPIN loss. Our results reveal a regulatory mode of E3 ligases in which the accessory E3 in LUBAC downregulates the main E3 by providing preferred substrates for autolinear ubiquitination. Thus, inhibition of HOIL-1L E3 represents a promising strategy for treating severe infections or immunodeficiency.


Assuntos
Proteínas de Transporte/fisiologia , Morte Celular , Doença Hepática Induzida por Substâncias e Drogas/imunologia , Dermatite de Contato/imunologia , Peptídeos e Proteínas de Sinalização Intracelular/fisiologia , Salmonelose Animal/imunologia , Ubiquitina-Proteína Ligases/fisiologia , Ubiquitina/metabolismo , Animais , Células Cultivadas , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Doença Hepática Induzida por Substâncias e Drogas/patologia , Dermatite de Contato/metabolismo , Dermatite de Contato/patologia , Embrião de Mamíferos/imunologia , Embrião de Mamíferos/metabolismo , Embrião de Mamíferos/patologia , Fibroblastos/imunologia , Fibroblastos/metabolismo , Fibroblastos/patologia , Regulação da Expressão Gênica , Camundongos Knockout , NF-kappa B/genética , NF-kappa B/metabolismo , Salmonella/patogenicidade , Salmonelose Animal/metabolismo , Salmonelose Animal/patologia , Índice de Gravidade de Doença , Transdução de Sinais , Ubiquitinação
8.
Metabolism ; 106: 154194, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32135161

RESUMO

BACKGROUND: Low-grade inflammation and metabolic dysregulation are common comorbidities of obesity, both of which are associated with alterations in iRhom2-regulated pro-inflammatory cytokine and epidermal growth factor receptor (EGFR) ligand signaling. OBJECTIVE: Our objective was to determine the role of iRhom2 in the regulation of low-grade inflammation and metabolic dysregulation in a murine model of diet-induced obesity. METHODS: Wild type (WT) and iRhom2-deficient mice were fed normal chow (NC) or a high-fat diet (HFD) starting at 5 weeks of age for up to 33 weeks. Body composition, glucose and insulin tolerance, feeding behavior, and indirect calorimetry were measured at defined time points. Adipose tissue cytokine expression and inflammatory lesions known as crown-like structures (CLS) were analyzed at the end-point of the study. RESULTS: iRhom2-deficient mice show accelerated fat gain on a HFD, accompanied by insulin resistance. Indirect calorimetry did not demonstrate changes in energy expenditure or food intake, but locomotor activity was significantly reduced in HFD iRhom2-deficient mice. Interestingly, CLS, macrophage infiltration, and tumor necrosis factor (TNF) production were decreased in adipose tissue from HFD iRhom2-deficient mice, but circulating cytokines were unchanged. In inguinal and perigonadal fat, the EGFR ligand amphiregulin was markedly induced in HFD controls but completely prevented in iRhom2-deficient mice, suggesting a potentially dominant role of EGFR-dependent mechanisms over TNF in the modulation of insulin sensitivity. CONCLUSIONS: This study elucidates a novel role for iRhom2 as an immuno-metabolic regulator that affects adipose tissue inflammation independent of insulin resistance.


Assuntos
Tecido Adiposo/metabolismo , Proteínas de Transporte/fisiologia , Dieta Hiperlipídica , Inflamação/patologia , Resistência à Insulina/genética , Obesidade/etiologia , Ganho de Peso/genética , Tecido Adiposo/patologia , Animais , Proteínas de Transporte/genética , Células Cultivadas , Dieta Hiperlipídica/efeitos adversos , Progressão da Doença , Regulação para Baixo/genética , Intolerância à Glucose/genética , Intolerância à Glucose/metabolismo , Intolerância à Glucose/patologia , Inflamação/genética , Inflamação/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Obesidade/genética , Obesidade/metabolismo , Obesidade/patologia , Paniculite/genética , Paniculite/metabolismo , Paniculite/patologia
9.
Anim Sci J ; 91(1): e13345, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32219949

RESUMO

During mammalian fertilization, sperm is fused with the oocyte's membrane, triggering the resumption of meiosis from the metaphase II arrest, the extrusion of the second polar body, and the exocytosis of cortical granules; these events are collectively called 'oocyte activation.' In all species studied to date, the transient rise in the cytosolic level of calcium (in particular, the repeated calcium increases called 'calcium oscillations' in mammals) is required for these events. Researchers have focused on identifying the factor(s) that can induce calcium oscillations during fertilization. Sperm-specific phospholipase C, i.e., PLC zeta (PLCζ), is a strong candidate of the factor(s), and several research groups using different species obtained evidence that PLCζ is a sperm factor that can induce calcium oscillations during fertilization. However, postacrosomal sheath Tryptophan-Tryptophan (WW)-domain-binding protein (PAWP) was recently shown to have a pivotal role in inducing calcium oscillations in some species. In this review, we focus on PLCζ and PAWP as sperm factors, and we discuss this controversy: Which of these two molecules survives as a sperm factor?


Assuntos
Proteínas de Transporte/fisiologia , Oócitos/fisiologia , Fosfolipases/fisiologia , Proteínas de Plasma Seminal/fisiologia , Interações Espermatozoide-Óvulo/fisiologia , Espermatozoides/fisiologia , Animais , Cálcio/metabolismo , Sinalização do Cálcio , Feminino , Fertilização/fisiologia , Masculino
10.
Science ; 367(6479): 763-768, 2020 02 14.
Artigo em Inglês | MEDLINE | ID: mdl-32054757

RESUMO

Effector-triggered immunity (ETI), induced by host immune receptors in response to microbial effectors, protects plants against virulent pathogens. However, a systematic study of ETI prevalence against species-wide pathogen diversity is lacking. We constructed the Pseudomonas syringae Type III Effector Compendium (PsyTEC) to reduce the pan-genome complexity of 5127 unique effector proteins, distributed among 70 families from 494 strains, to 529 representative alleles. We screened PsyTEC on the model plant Arabidopsis thaliana and identified 59 ETI-eliciting alleles (11.2%) from 19 families (27.1%), with orthologs distributed among 96.8% of P. syringae strains. We also identified two previously undescribed host immune receptors, including CAR1, which recognizes the conserved effectors AvrE and HopAA1, and found that 94.7% of strains harbor alleles predicted to be recognized by either CAR1 or ZAR1.


Assuntos
Arabidopsis/imunologia , Arabidopsis/microbiologia , Interações Hospedeiro-Patógeno/imunologia , Imunidade Inata/genética , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Imunidade Vegetal/genética , Pseudomonas syringae/patogenicidade , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/fisiologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/imunologia , Proteínas de Transporte/genética , Proteínas de Transporte/fisiologia , Genoma de Planta , Interações Hospedeiro-Patógeno/genética , Doenças das Plantas/genética , Pseudomonas syringae/genética
11.
J Virol ; 94(9)2020 04 16.
Artigo em Inglês | MEDLINE | ID: mdl-32102874

RESUMO

Flaviviruses, including dengue virus (DENV) and Zika virus (ZIKV), rely heavily on the availability of endoplasmic reticulum (ER) membranes throughout their life cycle, and degradation of ER membranes restricts flavivirus replication. Accordingly, DENV and ZIKV restrict ER turnover by protease-mediated cleavage of reticulophagy regulator 1 (RETREG1), also known as FAM134B, an autophagy receptor responsible for targeted ER sheet degradation. Given that the induction of autophagy may play an important role in flavivirus replication, the antiviral role of RETREG1 suggests that specialized autophagic pathways may have differential effects on the flavivirus life cycle. We previously identified BPI fold-containing family B member 3 (BPIFB3) as a regulator of autophagy that negatively controls enterovirus replication. Here, we show that in contrast to enteroviruses, BPIFB3 functions as a positive regulator of DENV and ZIKV infection and that its RNA interference-mediated silencing inhibits the formation of viral replication organelles. Mechanistically, we show that depletion of BPIFB3 enhances RETREG1-dependent reticulophagy, leading to enhanced ER turnover and the suppression of viral replication. Consistent with this, the antiviral effects of BPIFB3 depletion can be reversed by RETREG1 silencing, suggesting a specific role for BPIFB3 in regulating ER turnover. These studies define BPIFB3 as a required host factor for both DENV and ZIKV replication and further contribute to our understanding of the requirements for autophagy during flavivirus infection.IMPORTANCE Flaviviruses and other arthropod-transmitted viruses represent a widespread global health problem, with limited treatment options currently available. Thus, a better understanding of the cellular requirements for their infection is needed. Both DENV and ZIKV rely on expansion of the endoplasmic reticulum (ER) and the induction of autophagy to establish productive infections. However, little is known regarding the interplay between the requirements for autophagy initiation during infection and the mechanisms used by these viruses to avoid clearance through the autophagic pathway. Our study highlights the importance of the host factor BPIFB3 in regulating flavivirus replication and further confirms that the RETREG1-dependent reticulophagy pathway is antiviral to both DENV and ZIKV.


Assuntos
Proteínas de Transporte/metabolismo , Flavivirus/fisiologia , Replicação Viral/fisiologia , Autofagia , Proteínas de Transporte/fisiologia , Linhagem Celular , Vírus da Dengue/fisiologia , Retículo Endoplasmático/metabolismo , Retículo Endoplasmático/fisiologia , Retículo Endoplasmático/virologia , Flavivirus/metabolismo , Infecções por Flavivirus/virologia , Interações Hospedeiro-Patógeno/genética , Humanos , Interferência de RNA , Zika virus/fisiologia , Infecção por Zika virus/virologia
12.
Science ; 367(6476): 436-440, 2020 01 24.
Artigo em Inglês | MEDLINE | ID: mdl-31974253

RESUMO

The ability of the nervous system to sense cellular stress and coordinate protein homeostasis is essential for organismal health. Unfortunately, stress responses that mitigate disturbances in proteostasis, such as the unfolded protein response of the endoplasmic reticulum (UPRER), become defunct with age. In this work, we expressed the constitutively active UPRER transcription factor, XBP-1s, in a subset of astrocyte-like glia, which extended the life span in Caenorhabditis elegans Glial XBP-1s initiated a robust cell nonautonomous activation of the UPRER in distal cells and rendered animals more resistant to protein aggregation and chronic ER stress. Mutants deficient in neuropeptide processing and secretion suppressed glial cell nonautonomous induction of the UPRER and life-span extension. Thus, astrocyte-like glial cells play a role in regulating organismal ER stress resistance and longevity.


Assuntos
Caenorhabditis elegans/fisiologia , Estresse do Retículo Endoplasmático/fisiologia , Longevidade , Neuroglia/fisiologia , Neuropeptídeos/fisiologia , Animais , Caenorhabditis elegans/genética , Proteínas de Caenorhabditis elegans/genética , Proteínas de Caenorhabditis elegans/fisiologia , Proteínas de Transporte/genética , Proteínas de Transporte/fisiologia , Mutação , Agregados Proteicos/fisiologia , Transdução de Sinais , Fatores de Transcrição/genética , Fatores de Transcrição/fisiologia
13.
J Vasc Res ; 57(2): 76-85, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31968349

RESUMO

Diabetes mellitus (DM)-induced impairment of collateral formation has been demonstrated in subjects with coronary artery disease, which contributes to unfavorable prognosis among diabetic individuals. In our previous studies, thioredoxin1 (Trx1) activity was shown to be decreased in diabetic cardiac tissues, but the reason of Trx1 inactivation and whether it mediates the impaired angiogenesis in ischemic myocardium is still to be identified. As thioredoxin-interacting protein (TXNIP), an endogenous inhibitor of Trx, is overexpressed in DM due to carbohydrate response element within its promoter, we hypothesized that inhibition of Trx1 by enhanced TXNIP expression in endothelial cells may play a role in hyperglycemia-induced impairment of angiogenesis. In the present study, we found that high glucose-mediated increase of TXNIP expression and TXNIP-Trx1 interaction induced the impairment in endothelial cell function and survival, since these detrimental effects are rescued by silencing TXNIP with small interfering RNA. In diabetic mice, TXNIP knockdown or recombinant human Trx1 treatment counteracted the impairment of angiogenesis, alleviated myocardial ischemic injury, and improved survival rate. All these data implicate that TXNIP upregulation and subsequently the increased formation of TXNIP-Trx1 complex is a novel pathologic pathway by which DM induces insufficient angiogenesis and thereby exacerbates myocardial ischemia injury.


Assuntos
Diabetes Mellitus Experimental/complicações , Isquemia Miocárdica/fisiopatologia , Neovascularização Fisiológica/fisiologia , Tiorredoxinas/fisiologia , Animais , Proteínas de Transporte/fisiologia , Diabetes Mellitus Experimental/fisiopatologia , Células Endoteliais/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL
14.
Exp Cell Res ; 388(2): 111862, 2020 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-31982382

RESUMO

Thioredoxin-interacting protein (TXNIP), is identified as an inhibitor of the thiol oxidoreductase thioredoxin that acts endogenously, and is increased by high glucose (HG). In this study, we investigated the potential function of TXNIP on apoptosis of podocytes and its potential mechanism in vivo and in vitro in diabetic nephropathy (DN). TXNIP silencing attenuated HG-induced apoptosis and obliterated the activation of signaling pathways of mammalian target of rapamycin (mTOR) and p38 mitogen-activated protein kinase (MAPK) in conditionally immortalized mouse podocytes. Furthermore, the Raptor and Rictor shRNAs, mTOR specific inhibitor KU-0063794 and p38 MAPK inhibitor SB203580 were used to assess the role of mTOR or p38 MAPK pathway on podocyte apoptosis induced by HG. The Rictor and Raptor shRNAs and KU-0063794 appeared to reduce HG-induced apoptosis in podocytes. Simultaneously, SB203580 could also restrain HG-induced apoptosis in podocytes. Streptozotocin rendered equivalent diabetes in TXNIP-/- (TKO) and wild-type (WT) control mice. TXNIP deficiency mitigated renal injury in diabetic mice. Additionally, TXNIP deficiency also descended the apoptosis-related protein and Nox4 levels, the mTOR signaling activation and the p38 MAPK phosphorylation in podocytes of diabetic mice. All these data indicate that TXNIP deficiency may mitigate apoptosis of podocytes by inhibiting p38 MAPK or mTOR signaling pathway in DN, underlining TXNIP as a putative target for therapy.


Assuntos
Apoptose , Proteínas de Transporte/fisiologia , Nefropatias Diabéticas/prevenção & controle , Glucose/farmacologia , Podócitos/patologia , Serina-Treonina Quinases TOR/metabolismo , Tiorredoxinas/fisiologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Diabetes Mellitus Experimental/complicações , Nefropatias Diabéticas/etiologia , Nefropatias Diabéticas/metabolismo , Nefropatias Diabéticas/patologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fosforilação , Podócitos/metabolismo , Transdução de Sinais , Serina-Treonina Quinases TOR/genética , Proteínas Quinases p38 Ativadas por Mitógeno/genética
15.
Genes Cells ; 25(1): 65-70, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31721365

RESUMO

Atg2 is one of the essential factors for autophagy. Recent advance of structural and biochemical study on yeast Atg2 proposed that Atg2 tethers the edge of the isolation membrane (IM) to the endoplasmic reticulum and mediates direct lipid transfer (LT) from ER to IM for IM expansion. In mammals, two Atg2 orthologs, ATG2A and ATG2B, participate in autophagic process. Here we showed that human ATG2B possesses the membrane tethering (MT) and LT activity that was promoted by negatively charged membranes and an Atg18 ortholog WIPI4. By contrast, negatively charged membranes reduced the yeast Atg2 activities in the absence of Atg18. These results suggest that the MT/LT activity of Atg2 is evolutionally conserved although their regulation differs among species.


Assuntos
Proteínas Relacionadas à Autofagia/metabolismo , Proteínas de Transporte/metabolismo , Proteínas de Membrana/metabolismo , Proteínas de Transporte Vesicular/metabolismo , Autofagia , Proteínas Relacionadas à Autofagia/fisiologia , Transporte Biológico , Proteínas de Transporte/fisiologia , Retículo Endoplasmático/metabolismo , Humanos , Metabolismo dos Lipídeos/fisiologia , Lipídeos/fisiologia , Proteínas de Ligação a Fosfato/metabolismo , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Proteínas de Transporte Vesicular/fisiologia
16.
Mol Biol Cell ; 31(1): 59-79, 2020 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-31721635

RESUMO

Dense-core vesicles (DCVs) are secretory vesicles found in neurons and endocrine cells. DCVs package and release cargoes including neuropeptides, biogenic amines, and peptide hormones. We recently identified the endosome-associated recycling protein (EARP) complex and the EARP-interacting-protein EIPR-1 as proteins important for controlling levels of DCV cargoes in Caenorhabditis elegans neurons. Here we determine the role of mammalian EIPR1 in insulinoma cells. We find that in Eipr1 KO cells, there is reduced insulin secretion, and mature DCV cargoes such as insulin and carboxypeptidase E (CPE) accumulate near the trans-Golgi network and are not retained in mature DCVs in the cell periphery. In addition, we find that EIPR1 is required for the stability of the EARP complex subunits and for the localization of EARP and its association with membranes, but EIPR1 does not affect localization or function of the related Golgi-associated retrograde protein (GARP) complex. EARP is localized to two distinct compartments related to its function: an endosomal compartment and a DCV biogenesis-related compartment. We propose that EIPR1 functions with EARP to control both endocytic recycling and DCV maturation.


Assuntos
Proteínas de Caenorhabditis elegans/metabolismo , Proteínas de Transporte/metabolismo , Células Secretoras de Insulina/metabolismo , Vesículas Secretórias/fisiologia , Animais , Fenômenos Biofísicos , Caenorhabditis elegans/metabolismo , Proteínas de Caenorhabditis elegans/fisiologia , Proteínas de Transporte/fisiologia , Linhagem Celular , Endossomos/metabolismo , Complexo de Golgi/metabolismo , Neurônios/metabolismo , Neuropeptídeos/metabolismo , Transporte Proteico , Vesículas Secretórias/metabolismo , Proteínas de Transporte Vesicular/metabolismo , Rede trans-Golgi/metabolismo
17.
Inflamm Bowel Dis ; 26(2): 242-253, 2020 01 06.
Artigo em Inglês | MEDLINE | ID: mdl-31586441

RESUMO

BACKGROUND: Tumor necrosis factor (TNF)-α is a major proinflammatory cytokine that plays a key role in inflammatory bowel disease (IBD). Inactive rhomboid protein 2 (iRhom2) is essential for activating TNF-α-converting enzyme (TACE) in immune cells, which regulates TNF-α release. The aim of the study was to investigate the role of iRhom2 in intestinal inflammation in IBD. METHODS: The expression of iRhom2 and TACE in lipopolysaccharide (LPS)-stimulated COLO 205 and RAW 264.7 cells was assessed by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis. The expression of iRhom2 and TACE in the colonic tissue of IBD patients and 2,4,6-trinitrobenzenesulfonic acid solution (TNBS)-treated mice was determined by RT-PCR and immunohistochemistry. To assess the role of iRhom2 in intestinal inflammation, colitis was induced in wild-type and iRhom2-/- mice by the administration of TNBS enema. RESULTS: In LPS-stimulated COLO 205 and RAW 264.7 cells, the mRNA and protein levels of TACE and iRhom2 were upregulated. The expression of TACE and iRhom2 in the colon of the IBD patients and TNBS-treated mice was significantly enhanced. The inflammatory cells that expressed high levels of iRhom2 in the colon were identified as macrophages. Finally, iRhom2 deficiency ameliorated TNBS-induced colitis by inhibiting TNF-α release. CONCLUSIONS: iRhom2 has an important role in intestinal inflammation through TNF-α secretion in immune cells, which suggests that iRhom2 could be a novel therapeutic target for IBD.


Assuntos
Proteínas de Transporte/fisiologia , Colite/etiologia , Inflamação/etiologia , Intestinos/imunologia , Macrófagos/imunologia , Fator de Necrose Tumoral alfa/metabolismo , Animais , Colite/patologia , Citocinas , Inflamação/metabolismo , Inflamação/patologia , Intestinos/efeitos dos fármacos , Intestinos/patologia , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Macrófagos/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Estresse Oxidativo , Ácido Trinitrobenzenossulfônico/toxicidade
18.
Nat Prod Res ; 34(6): 830-837, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30445855

RESUMO

Amphibian egg-jelly coat plays an important role in successful fertilization and development. Here, we ask whether proteins like allurin in the jelly coats of frog eggs might influence fertilization rate success. Using in vitro fertilization of Albanian water frog, Pelophylax shqipericus, we found that body cavity eggs or eggs deprived of jelly coat were not fertilized, compromising the success of in vitro fertilization procedure. When de-jellied eggs were inseminated with sperm suspension, the fertilization efficiency is dramatically decreased even inhibited, suggesting that the gel structure is one of the major factors in the achievement of fertilization in the frogs. Fertilization of de-jellied eggs with sperm pre-treated with egg jelly coat, restored the fertilization competency. Such a result suggests that egg jelly coat probably guides the sperm to the egg surface while maintaining the fertilization ability, contributing to a successful in vitro fertilization of Pelophylax shqipericus.


Assuntos
Proteínas de Transporte/fisiologia , Proteínas do Ovo/fisiologia , Espécies em Perigo de Extinção , Fertilização In Vitro/métodos , Ranidae/fisiologia , Animais , Feminino , Fertilização , Masculino , Óvulo , Espermatozoides , Água
19.
Plant Biotechnol J ; 18(3): 756-769, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31469486

RESUMO

Nonspecific lipid transfer proteins (nsLTPs) play critical roles in plant development and response to abiotic stresses. Here, we found that a rice lipid transfer protein, OsLTPL159, was associated with cold tolerance at the early seedling stage. Overexpression of an OsLTPL159IL 112 allele from the cold-tolerant introgression line IL112 in either the japonica variety Zhonghua17 (ZH17) or the indica variety Teqing background dramatically enhanced cold tolerance. In addition, down-regulation of the expression of OsLTPL159 in the japonica variety ZH17 by RNA interference (RNAi) significantly decreased cold tolerance. Further transcriptomic, physiological and histological analysis showed that the OsLTPL159IL 112 allele likely enhanced the cold tolerance of rice at the early seedling stage by decreasing the toxic effect of reactive oxygen species, enhancing cellulose deposition in the cell wall and promoting osmolyte accumulation, thereby maintaining the integrity of the chloroplasts. Notably, overexpression of another allele, OsLTPL159GC 2 , from the recipient parent Guichao 2 (GC2), an indica variety, did not improve cold tolerance, indicating that the variations in the OsLTPL159 coding region of GC2 might disrupt its function for cold tolerance. Further sequence comparison found that all 22 japonica varieties surveyed had an OsLTPL159 haplotype identical to IL112 and were more cold-tolerant than the surveyed indica varieties, implying that the variations in OsLTPL159 might be associated with differential cold tolerance of japonica and indica rice. Therefore, our findings suggest that the OsLTPL159 allele of japonica rice could be used to improve cold tolerance of indica rice through a molecular breeding strategy.


Assuntos
Proteínas de Transporte/fisiologia , Temperatura Baixa , Oryza/fisiologia , Proteínas de Plantas/fisiologia , Estresse Fisiológico , Perfilação da Expressão Gênica , Plântula/fisiologia
20.
Sci Rep ; 9(1): 18712, 2019 12 10.
Artigo em Inglês | MEDLINE | ID: mdl-31822696

RESUMO

The E. coli membrane protein ZipA, binds to the tubulin homologue FtsZ, in the early stage of cell division. We isolated ZipA in a Styrene Maleic Acid lipid particle (SMALP) preserving its position and integrity with native E. coli membrane lipids. Direct binding of ZipA to FtsZ is demonstrated, including FtsZ fibre bundles decorated with ZipA. Using Cryo-Electron Microscopy, small-angle X-ray and neutron scattering, we determine the encapsulated-ZipA structure in isolation, and in complex with FtsZ to a resolution of 1.6 nm. Three regions can be identified from the structure which correspond to, SMALP encapsulated membrane and ZipA transmembrane helix, a separate short compact tether, and ZipA globular head which binds FtsZ. The complex extends 12 nm from the membrane in a compact structure, supported by mesoscale modelling techniques, measuring the movement and stiffness of the regions within ZipA provides molecular scale analysis and visualisation of the early divisome.


Assuntos
Proteínas de Bactérias/metabolismo , Proteínas de Transporte/metabolismo , Proteínas de Ciclo Celular/metabolismo , Divisão Celular/fisiologia , Proteínas do Citoesqueleto/metabolismo , Proteínas de Escherichia coli/metabolismo , Proteínas de Bactérias/fisiologia , Proteínas de Transporte/fisiologia , Proteínas de Transporte/ultraestrutura , Proteínas de Ciclo Celular/fisiologia , Proteínas de Ciclo Celular/ultraestrutura , Microscopia Crioeletrônica/métodos , Proteínas do Citoesqueleto/fisiologia , Escherichia coli/metabolismo , Proteínas de Escherichia coli/fisiologia , Proteínas de Escherichia coli/ultraestrutura , Proteínas de Membrana/metabolismo , Ligação Proteica
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