scholarly journals Grouper TRAF4, a Novel, CP-Interacting Protein That Promotes Red-Spotted Grouper Nervous Necrosis Virus Replication

2021 ◽  
Vol 22 (11) ◽  
pp. 6136
Author(s):  
Siting Wu ◽  
Mengshi Sun ◽  
Xin Zhang ◽  
Jiaming Liao ◽  
Mengke Liu ◽  
...  
Keyword(s):  
Tumor Necrosis Factor Receptor ◽  
Nuclear Factor Κb ◽  
Inflammatory Factors ◽  
Nervous Necrosis Virus ◽  
Biological Processes ◽  
Necrosis Virus ◽  
Interacting Protein ◽  
Acid Protein ◽  
Amino Acid Protein

Tumor necrosis factor receptor-associated factors (TRAFs) play important roles in the biological processes of immune regulation, the inflammatory response, and apoptosis. TRAF4 belongs to the TRAF family and plays a major role in many biological processes. Compared with other TRAF proteins, the functions of TRAF4 in teleosts have been largely unknown. In the present study, the TRAF4 homologue (EcTRAF4) of the orange-spotted grouper was characterized. EcTRAF4 consisted of 1413 bp encoding a 471-amino-acid protein, and the predicted molecular mass was 54.27 kDa. EcTRAF4 shares 99.79% of its identity with TRAF4 of the giant grouper (E. lanceolatus). EcTRAF4 transcripts were ubiquitously and differentially expressed in all the examined tissues. EcTRAF4 expression in GS cells was significantly upregulated after stimulation with red-spotted grouper nervous necrosis virus (RGNNV). EcTRAF4 protein was distributed in the cytoplasm of GS cells. Overexpressed EcTRAF4 promoted RGNNV replication during viral infection in vitro. Yeast two-hybrid and coimmunoprecipitation assays showed that EcTRAF4 interacted with the coat protein (CP) of RGNNV. EcTRAF4 inhibited the activation of IFN3, IFN-stimulated response element (ISRE), and nuclear factor-κB (NF-κB). Overexpressed EcTRAF4 also reduced the expression of interferon (IFN)-related molecules and pro-inflammatory factors. Together, these results demonstrate that EcTRAF4 plays crucial roles in RGNNV infection.

scholarly journals LASS3 (longevity assurance homologue 3) is a mainly testis-specific (dihydro)ceramide synthase with relatively broad substrate specificity

2006 ◽  
Vol 398 (3) ◽  
pp. 531-538 ◽  
Author(s):  
Yukiko Mizutani ◽  
Akio Kihara ◽  
Yasuyuki Igarashi
Keyword(s):  
Amino Acid ◽  
Substrate Specificity ◽  
Family Members ◽  
Fatty Acyl ◽  
Ceramide Synthase ◽  
Broad Substrate Specificity ◽  
Acid Protein ◽  
Amino Acid Protein

The LASS (longevity assurance homologue) family members are highly conserved from yeasts to mammals. Five mouse and human LASS family members, namely LASS1, LASS2, LASS4, LASS5 and LASS6, have been identified and characterized. In the present study we cloned two transcriptional variants of hitherto-uncharacterized mouse LASS3 cDNA, which encode a 384-amino-acid protein (LASS3) and a 419-amino-acid protein (LASS3-long). In vivo, [3H]dihydrosphingosine labelling and electrospray-ionization MS revealed that overproduction of either LASS3 isoform results in increases in several ceramide species, with some preference toward those having middle- to long-chain-fatty acyl-CoAs. A similar substrate preference was observed in an in vitro (dihydro)ceramide synthase assay. These results indicate that LASS3 possesses (dihydro)ceramide synthesis activity with relatively broad substrate specificity. We also found that, except for a weak display in skin, LASS3 mRNA expression is limited almost solely to testis, implying that LASS3 plays an important role in this gland.

In vitro neutralization of nervous necrosis virus by a nanobody binding to the protrusion domain of capsid protein

Aquaculture ◽  
2021 ◽  
pp. 737654
Author(s):  
Song Zhu ◽  
Bo Miao ◽  
Yu-Zhou Zhang ◽  
Wei-Wei Zeng ◽  
De-Shou Wang ◽  
...  
Keyword(s):  
Capsid Protein ◽  
Nervous Necrosis Virus ◽  
Necrosis Virus

scholarly journals TAB1 regulates glycolysis and activation of macrophages in diabetic nephropathy

2020 ◽  
Vol 69 (12) ◽  
pp. 1215-1234
Author(s):  
Hanxu Zeng ◽  
Xiangming Qi ◽  
Xingxin Xu ◽  
Yonggui Wu
Keyword(s):  
Diabetic Nephropathy ◽  
Transforming Growth Factor ◽  
Lentiviral Vector ◽  
Hypoxia Inducible Factor ◽  
Damage Index ◽  
Transforming Growth Factor Β ◽  
Nuclear Factor Κb ◽  
Inflammatory Factors

Abstract Objective and design Macrophages exhibit strong phenotypic plasticity and can mediate renal inflammation by polarizing into an M1 phenotype. They play a pivotal role in diabetic nephropathy (DN). Here, we have investigated the regulatory role of transforming growth factor β-activated kinase 1-binding protein 1 (TAB1) in glycolysis and activation of macrophages during DN. Methods TAB1 was inhibited using siRNA in high glucose (HG)-stimulated bone marrow-derived macrophages (BMMs) and lentiviral vector-mediated TAB1 knockdown was used in streptozotocin (STZ)-induced diabetic mice. Western blotting, flow cytometry, qRT-PCR, ELISA, PAS staining and immunohistochemical staining were used for assessment of TAB1/nuclear factor-κB (NF-κB)/hypoxia-inducible factor-1α (HIF-1α), iNOS, glycolysis, inflammation and the clinical and pathological manifestations of diabetic nephropathy. Results We found that TAB1/NF-κB/HIF-1α, iNOS and glycolysis were up-regulated in BMMs under HG conditions, leading to release of further inflammatory factors, Downregulation of TAB1 could inhibit glycolysis/polarization of macrophages and inflammation in vivo and in vitro. Furthermore, albuminuria, the tubulointerstitial damage index and glomerular mesangial expansion index of STZ-induced diabetic nephropathy mice were decreased by TAB1 knockdown. Conclusions Our results suggest that the TAB1/NF-κB/HIF-1α signaling pathway regulates glycolysis and activation of macrophages in DN.

scholarly journals SVIP Is a Novel VCP/p97-interacting Protein Whose Expression Causes Cell Vacuolation

2003 ◽  
Vol 14 (1) ◽  
pp. 262-273 ◽  
Author(s):  
Masami Nagahama ◽  
Mie Suzuki ◽  
Yuko Hamada ◽  
Kiyotaka Hatsuzawa ◽  
Katsuko Tani ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Coiled Coil ◽  
Adaptor Proteins ◽  
Interacting Protein ◽  
Cellular Processes ◽  
Coiled Coil Region ◽  
Acid Protein ◽  
Extensive Cell ◽  
Dependent Protein ◽  
Amino Acid Protein

VCP/p97 is involved in a variety of cellular processes, including membrane fusion and ubiquitin-dependent protein degradation. It has been suggested that adaptor proteins such as p47 and Ufd1p confer functional versatility to VCP/p97. To identify novel adaptors, we searched for proteins that interact specifically with VCP/p97 by using the yeast two-hybrid system, and discovered a novel VCP/p97-interacting protein named smallVCP/p97-interactingprotein (SVIP). Rat SVIP is a 76-amino acid protein that contains two putative coiled-coil regions, and potential myristoylation and palmitoylation sites at the N terminus. Binding experiments revealed that the N-terminal coiled-coil region of SVIP, and the N-terminal and subsequent ATP-binding regions (ND1 domain) of VCP/p97, interact with each other. SVIP and previously identified adaptors p47 and ufd1p interact with VCP/p97 in a mutually exclusive manner. Overexpression of full-length SVIP or a truncated mutant did not markedly affect the structure of the Golgi apparatus, but caused extensive cell vacuolation reminiscent of that seen upon the expression of VCP/p97 mutants or polyglutamine proteins in neuronal cells. The vacuoles seemed to be derived from endoplasmic reticulum membranes. These results together suggest that SVIP is a novelVCP/p97 adaptor whose function is related to the integrity of the endoplasmic reticulum.

Bacterial lipopolysaccharide directly induces angiogenesis through TRAF6-mediated activation of NF-κB and c-Jun N-terminal kinase

Blood ◽  
2003 ◽  
Vol 102 (5) ◽  
pp. 1740-1742 ◽  
Author(s):  
Ingrid Pollet ◽  
Christy J. Opina ◽  
Carla Zimmerman ◽  
Kevin G. Leong ◽  
Fred Wong ◽  
...  
Keyword(s):  
Intracellular Signaling ◽  
Tumor Necrosis Factor Receptor ◽  
Nuclear Factor Κb ◽  
Dominant Negative ◽  
Bacterial Lipopolysaccharide ◽  
Intracellular Signaling Pathway ◽  
Necrosis Factor

AbstractThe intracellular pathways by which inflammatory mediators transmit their angiogenic signals is not well studied. The effects of a potent inflammatory mediator, bacterial lipopolysaccharide (LPS), are transmitted through Toll-like receptors (TLRs). A major, although not exclusive, LPS/TLR intracellular signaling pathway is routed through TNF (tumor necrosis factor) receptor associated factor 6 (TRAF6). In this report we demonstrate that LPS directly stimulates endothelial sprouting in vitro. By blocking TRAF6 activity using retroviral expression of a dominant-negative TRAF6 in endothelial cells, we show that TRAF6 is absolutely required for the LPS-initiated angiogenic response in vitro and in vivo. Inhibition of either c-Jun N-terminal kinase (JNK) activity or nuclear factor κB (NF-κB) activity, downstream of TRAF6, is sufficient to inhibit LPS-induced endothelial sprouting. In contrast, only inhibition of NF-κB, but not JNK, activity blocks basic fibroblast growth factor (bFGF)–induced angiogenesis. Our findings thus demonstrate a direct endothelial-stimulatory role of LPS in initiating angiogenesis through activation of TRAF6-dependent signaling pathways.

scholarly journals TBK1 inhibitor exerts anti-proliferative effect on glioblastoma multiforme cells

2021 ◽  
Author(s):  
Sarah A Scuderi ◽  
Marika Lanza ◽  
Giovanna Casili ◽  
Francesca Esposito ◽  
Cristina Colarossi ◽  
...  
Keyword(s):  
Glioblastoma Multiforme ◽  
Cell Viability ◽  
Ex Vivo ◽  
Tumor Necrosis Factor Receptor ◽  
Nuclear Factor Κb ◽  
Potential Effect ◽  
Ex Vivo Model ◽  
Iκb Kinase ◽  
Pathway Activation

Glioma are common malignant brain tumors, among which glioblastoma multiforme (GBM) has the worst prognosis. Different studies of GBM revealed that targeting nuclear-factor-κB (NF-κB) induced an attenuation tumor proliferation and prolonged cell survival. TBK1 (TANK {TRAF (TNF (tumor-necrosis-factor) receptor-associated factor)-associated NF-κB activator}-binding kinase 1) is a serine/threonine-protein kinase and it is a member of the IκB kinase (IKK) family, involved in NF-κB pathway activation. The aim of this study was to investigate the potential effect of BX795, an inhibitor of TBK1, in an in vitro and ex vivo model of GBM. GBM cell lines (U87 and U138) and primary GBM cells were treated with different concentrations of BX795 at different time-points (24, 48 and 72h) to evaluate cell viability, autophagy, inflammation and apoptosis. Our results demonstrated that BX795 10 μM was able to reduce cell viability, showing antiproliferative effect both in U87, U138 and primary GBM cells. Moreover, treatment with BX795 10 μM increased the pro-apoptotic proteins Bax, p53, caspase-3 and caspase-9 whereas the anti-apoptotic Bcl-2 expression was reduced. Additionally our results showed a marked decrease of autophagy following BX795 treatment, reducing Atg7, Atg 5/12 and AKT expression. The anti-inflammatory effect of BX795 was demonstrated by a significantly reduction of NIK, IKKα and TNF-α expression, accompanied by a down-regulation of angiogenesis. Furthermore, in primary GBMs cell, BX795 10 μM was able to reduce TBK1 pathway activation and SOX3 expression. In conclusion, these findings showed that TBK1 is involved in GBM proliferation demonstrating that the inhibitor BX795, thanks its abilities, could improve therapeutic strategies for GBM treatment.

scholarly journals Grtp1, A Novel Gene Regulated by Growth Hormone

Endocrinology ◽  
2001 ◽  
Vol 142 (10) ◽  
pp. 4568-4571 ◽  
Author(s):  
Chunxia Lu ◽  
John Kasik ◽  
Dietrich A. Stephan ◽  
Shaohua Yang ◽  
Mark A. Sperling ◽  
...  
Keyword(s):  
Northern Blot Analysis ◽  
Small Gtpases ◽  
Chromosome 8 ◽  
Taqman Assay ◽  
Activator Proteins ◽  
Novel Gene ◽  
Acid Protein ◽  
Vitro Model ◽  
Amino Acid Protein

Abstract An in vitro model of GH-responsive cells was subjected to microarray analysis to identify a novel gene regulated by GH. This 258 amino acid protein, we term GH Regulated TBC Protein-1 (GRTP1), contains the TBC signature motif of GTPase activator proteins of Rab-like small GTPases. Northern blot analysis revealed a 1.3 kb major mRNA species, most abundant in testes. TaqMan assay confirmed that in the mouse, Grtp1 is expressed at highest levels in testes, with lesser abundance in intestine, kidney, lung, and liver. In the testis, expression of Grtp1 significantly increases post-pubertally. Administration of GH to mice increased levels of GRTP1 mRNA in testes (140%), but decreased GRTP1 mRNA abundance in kidney (50%) and liver (25%). Grtp1 was localized to mouse proximal chromosome 8. Orthologs of this protein are present in human, mouse, rat, and drosophila suggesting that GRTP1 has an important biological role(s).

scholarly journals Establishment of an infectious RNA transcription system for Striped jack nervous necrosis virus, the type species of the betanodaviruses

2001 ◽  
Vol 82 (11) ◽  
pp. 2653-2662 ◽  
Author(s):  
Tokinori Iwamoto ◽  
Kazuyuki Mise ◽  
Koh-ichiro Mori ◽  
Misao Arimoto ◽  
Toshihiro Nakai ◽  
...  
Keyword(s):  
Type Species ◽  
Fish Cell ◽  
Cdna Clones ◽  
Nervous Necrosis Virus ◽  
Necrosis Virus ◽  
T7 Promoter ◽  
Genomic Rnas ◽  
Transcription System ◽  
Indirect Immunofluorescence Staining

A system has been established to produce infectious RNA transcripts for Striped jack nervous necrosis virus (SJNNV), the type species of the betanodaviruses, which infect fish. An enzymological analysis suggested that both RNA1 and RNA2 of SJNNV have a 5′ cap. Both RNAs were largely resistant to 3′ polyadenylation and ligation, suggesting the presence of an interfering 3′ structure, while a small quantity of viral RNAs were polyadenylated in vitro. The complete 5′ and 3′ non-coding sequences of both segments were determined using the rapid amplification of cDNA ends method. Based on the terminal sequences obtained, RT–PCR was carried out and plasmid clones containing full-length cDNA copies of both RNAs, positioned downstream of a T7 promoter, were constructed. These plasmids were cleaved at a unique restriction site just downstream of the 3′ terminus of each SJNNV sequence and were transcribed in vitro into RNA with a cap structure analogue. A mixture of the transcripts was transfected into the fish cell line E-11. Using indirect immunofluorescence staining with anti-SJNNV serum, fluorescence was observed specifically in these transfected cells; this culture supernatant exhibited pathogenicity to striped jack larvae. Northern blot analysis of E-11 cells infected with the recombinant virus or SJNNV showed small RNA (ca. 0·4 kb) that was newly synthesized and corresponded to the 3′-terminal region of RNA1. Finally, the complete nucleotide sequences of these functional cDNAs (RNA1, 3107 nt; RNA2, 1421 nt) were determined. This is the first report of betanodavirus cDNA clones from which infectious genomic RNAs can be transcribed.

CrkRS

2001 ◽  
Vol 114 (14) ◽  
pp. 2591-2603 ◽  
Author(s):  
Tun K. Ko ◽  
Emma Kelly ◽  
Jonathon Pines
Keyword(s):  
Protein Kinase ◽  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Genetic Locus ◽  
Kinase Domain ◽  
Speckled Pattern ◽  
Protein Kinase Domain ◽  
Acid Protein ◽  
Amino Acid Protein

We have isolated and characterised a novel human protein kinase, Cdc2-related kinase with an arginine/serine-rich (RS) domain (CrkRS), that is most closely related to the cyclin-dependent kinase (CDK) family. CrkRS is a 1490 amino acid protein, the largest CDK-related kinase so far isolated. The protein kinase domain of CrkRS is 89% identical to the 46 kDa CHED protein kinase, but outside the kinase domains the two proteins are completely unrelated. CrkRS has extensive proline-rich regions that match the consensus for SH3 and WW domain binding sites, and an RS domain that is predominantly found in splicing factors. CrkRS is ubiquitously expressed in tissues, and maps to a single genetic locus. There are closely related protein kinases in both the Drosophila and Caenorhabditis elegans genomes. Consistent with the presence of an RS domain, anti-CrkRS antibodies stain nuclei in a speckled pattern, overlapping with spliceosome components and the hyperphosphorylated form of RNA polymerase II. Like RNA polymerase II, CrkRS is a constitutive MPM-2 antigen throughout the cell cycle. Anti-CrkRS immunoprecipitates phosphorylate the C-terminal domain of RNA polymerase II in vitro. Thus CrkRS may be a novel, conserved link between the transcription and splicing machinery.

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