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Ernst G. Malygin - One of the best experts on this subject based on the ideXlab platform.
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Identification of the gene encoding vaccinia virus immunodominant Protein P35
Gene, 1994Co-Authors: Victor V. Zinoviev, Nicolay A. Tchikaev, Oleg Yu. Chertov, Ernst G. MalyginAbstract:Abstract The major envelope Protein, P35, of vaccinia virus (VV; strain LIVP) was purified by extraction from virions with the non-ionic detergent Nonidet P-40. The Protein was cleaved with CNBr. Four homogeneous peptides were isolated and their N-terminal amino-acid (aa) sequences determined. A computer search of a Protein sequence databank revealed complete identity of the determined sequences with aa 44-63, 144-149, 154-165 and 224-238 of ORF H3 of the HindIII-H fragment of the VV genome [Rosel et al J. Virol. 60 (1989) 436-446]. Earlier, Gordon et al. [Virology 167 (1988) 361-369] determined that the P35 surface Protein of VV strain IHD-W is encoded by the H6 gene. Muravlev et al. [Biopolymery i kletka 6 (1990) 83-89 (Russian)] deduced from their data that gene A2 encodes this prominent antigen. Taking into account this ambiguity, we cloned the genes H3, H6 and A2 in expression vectors, prepared the specific antisera against the expression products and conducted the immunochemical analysis of the recombinant and native VV-specific Proteins. It has been established that the H6 codes for an early Protein that is found only in the infected cell extracts, but is absent in mature virions. The immunodominant Protein P35 of VV strain LIVP is encoded by the gene H3. The gene A2 Protein product is present mainly in the infected cell extract, but the antiserum against the A2 product shows a rather weak interaction with the 35-kDa fraction of structural VV Proteins resolved by electrophoresis.
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Identification of the gene for the immunodominant P35 Protein from vaccinia virus
Molekuliarnaia biologiia, 1992Co-Authors: L. G. Ovechkina, Ernst G. Malygin, Liudmila Matskova, S M Balakhnin, Chertov OiuAbstract:A major immunodominant envelope Protein P35 of vaccina virus was purified by means of extraction from virions with detergent NP-40. The Protein was cleaved with CNBr, four homogenous peptides were isolated and their N-terminal amino acid sequences were determined. Computer search in a Protein sequences data bank revealed that the immunodominant Protein P35 of vaccinia virus is encoded by H3 gene in HindIII-H fragment of vaccinia virus genome.
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Amino acid sequence determination of vaccinia virus immunodominant Protein P35 and identification of the gene.
Biomedical science, 1991Co-Authors: Chertov Oyu, L. G. Ovechkina, Ernst G. MalyginAbstract:A major immunodominant envelope Protein of vaccinia virus (Protein P35) was purified by extraction from virions with the nonionic detergent Nonidet P-40. The Protein was cleaved with cyanogen bromide. Four homogeneous peptides were isolated and their N-terminal amino acid sequences determined. A computer search of a Protein-sequence data bank revealed complete identity of the determined sequences with sequences 44-63, 144-149, 154-165, and 224-238 of ORF H3 of the HindIII-H fragment of the vaccinia virus genome (Rosel et al 1986). It has therefore been established that the immunodominant Protein P35 of vaccinia virus is encoded by the gene in the HindIII-H fragment of the vaccinia virus genome.
Balraj Singh - One of the best experts on this subject based on the ideXlab platform.
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Evidence for an important role of serine 16 and its phosphorylation in the stabilization of c-Mos
Oncogene, 1999Co-Authors: Chau D Pham, Vijayalakshmi B Vuyyuru, Yandan Yang, Balraj SinghAbstract:The c-Mos serine/threonine Protein kinase is an essential component of cytostatic factor (CSF), which is required for metaphase II arrest of eggs in vertebrates. Previously, we showed that c-Mos residue Ser-16 is phosphorylated in the ts110 Mo-MuSV-encoded Gag-Mos fusion Protein. Here we provide evidence that Mos is phosphorylated at Ser-16 in transfected COS-1 cells. To investigate the role of this phosphorylation, Ser-16 was substituted with alanine or glutamic acid in full-length v-Mos (an Env-Mos fusion Protein that contains 31 additional amino acids at the amino terminus of c-Mos), its mouse c-Mos equivalent version (v-Mos residues 32-374, hereafter referred to as Mos), and mouse c-Mos. Constructs expressing mutant versions of Mos were transfected into COS-1 and NIH3T3 cells in a transient and stable manner, respectively. Synthesis and proteolysis of Mos were evaluated by pulse-chase analysis of ^35S-methionine-labeled Proteins. Our findings indicate that the S16A mutant of Mos was highly unstable. It accumulated to approximately 10% of the level of wild-type Mos or its S16E mutant. In addition, the S16A mutation but not the S16E mutation inhibited Mos interaction with a cellular Protein, P35, suggesting that phosphorylation at Ser-16 may promote Mos interaction with P35. As expected from its destabilizing effect, the S16A mutation caused a dramatic decrease in the cellular transforming activity of Mos (determined by soft-agar colony-formation assay with the stably transfected NIH3T3 cells), which is known to correlate with its CSF function. Efficient ubiquitin-mediated proteolysis of c-Mos requires proline as the second residue from the amino-terminus. In contrast to Mos, neither the stability nor Protein kinase activity of v-Mos (in which c-Mos residue Pro-2 becomes Pro-33) was affected by the S16A mutation. To provide further proof that, similar to c-Mos, the S16A mutant is recognized by the proteolysis system through Pro-2, we show that the effect of the S16A mutation is reversed by the Pro-2-Ala mutation. Thus, our results indicate that Ser-16 has an important role in the regulation of c-Mos and that phosphorylation at Ser-16 may inhibit proteolysis of c-Mos.
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Evidence for an important role of serine 16 and its phosphorylation in the stabilization of c-Mos
Oncogene, 1999Co-Authors: Chau D Pham, Vijayalakshmi B Vuyyuru, Yandan Yang, Wenlong Bai, Balraj SinghAbstract:The c-Mos serine/threonine Protein kinase is an essential component of cytostatic factor (CSF), which is required for metaphase II arrest of eggs in vertebrates. Previously, we showed that c-Mos residue Ser-16 is phosphorylated in the ts110 Mo-MuSV-encoded Gag-Mos fusion Protein. Here we provide evidence that Mos is phosphorylated at Ser-16 in transfected COS-1 cells. To investigate the role of this phosphorylation, Ser-16 was substituted with alanine or glutamic acid in full-length v-Mos (an Env-Mos fusion Protein that contains 31 additional amino acids at the amino terminus of c-Mos), its mouse c-Mos equivalent version (v-Mos residues 32-374, hereafter referred to as Mos), and mouse c-Mos. Constructs expressing mutant versions of Mos were transfected into COS-1 and NIH3T3 cells in a transient and stable manner, respectively. Synthesis and proteolysis of Mos were evaluated by pulse-chase analysis of ^35S-methionine-labeled Proteins. Our findings indicate that the S16A mutant of Mos was highly unstable. It accumulated to approximately 10% of the level of wild-type Mos or its S16E mutant. In addition, the S16A mutation but not the S16E mutation inhibited Mos interaction with a cellular Protein, P35, suggesting that phosphorylation at Ser-16 may promote Mos interaction with P35. As expected from its destabilizing effect, the S16A mutation caused a dramatic decrease in the cellular transforming activity of Mos (determined by soft-agar colony-formation assay with the stably transfected NIH3T3 cells), which is known to correlate with its CSF function. Efficient ubiquitin-mediated proteolysis of c-Mos requires proline as the second residue from the amino-terminus. In contrast to Mos, neither the stability nor Protein kinase activity of v-Mos (in which c-Mos residue Pro-2 becomes Pro-33) was affected by the S16A mutation. To provide further proof that, similar to c-Mos, the S16A mutant is recognized by the proteolysis system through Pro-2, we show that the effect of the S16A mutation is reversed by the Pro-2-Ala mutation. Thus, our results indicate that Ser-16 has an important role in the regulation of c-Mos and that phosphorylation at Ser-16 may inhibit proteolysis of c-Mos.
Rick T Dobrowsky - One of the best experts on this subject based on the ideXlab platform.
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Role of a cdk5-associated Protein, P35, in herpes simplex virus type 1 replicationin vivo
Journal of NeuroVirology, 2010Co-Authors: Steve D Haenchen, Jeff A Utter, Adam M Bayless, Rick T Dobrowsky, David J. DavidoAbstract:Previous studies have shown that herpes simplex virus type 1 (HSV-1) replication is inhibited by the cyclin-dependent kinase (cdk) inhibitor roscovitine. One roscovitine-sensitive cdk that functions in neurons is cdk5, which is activated in part by its binding partner, P35. Because HSV establishes latent infections in sensory neurons, we sought to determine the role P35 plays in HSV-1 replication in vivo . For these studies, wild-type (wt) and P35-/- mice were infected with HSV-1 using the mouse ocular model of HSV latency and reactivation. The current results indicate that P35 is an important determinant of viral replication in vivo .
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P35/Cyclin-dependent kinase 5 is required for protection against β-amyloid-induced cell death but not tau phosphorylation by ceramide
Journal of Molecular Neuroscience, 2007Co-Authors: Kathleen I. Seyb, Sabah Ansar, Jennifer Bean, Mary L. Michaelis, Rick T DobrowskyAbstract:Ceramide is a bioactive sphingolipid, that can prevent calpain activation and β-amyloid (Aß) neurotoxicity in cortical neurons. Recent evidence supports Aß induction of a calpain-dependent cleavage of the cyclin-dependent kinase 5 (cdk5) regulatory Protein P35 that contributes to tau hyperphosphorylation and neuronal death. Using cortical neurons isolated from wild-type and P35 knockout mice, we investigated whether ceramide required P35/cdk5 to protect against Aß-induced cell death and tau phosphorylation. Ceramide inhibited Aß-induced calpain activation and cdk5 activity in wild-type neurons and protected against neuronal death and tau hyperphosphorylation. Interestingly, Aß also increased cdk5 activity in P35^−/− neurons, suggesting that the alternate cdk5 regulatory Protein, p39, might mediate this effect. In P35 null neurons, ceramide blocked Aß-induced calpain activation but did not inhibit cdk5, activity or cell death. However, ceramide blocked tau hyperphosphorylation potentially via inhibition of glycogen synthase kinase-3β. These data suggest that ceramide can regulate Aß cell toxicity in a P35/cdk5-dependent manner.
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P35/Cyclin-dependent kinase 5 is required for protection against β-amyloid-induced cell death but not tau phosphorylation by ceramide
Journal of Molecular Neuroscience, 2007Co-Authors: Kathleen I. Seyb, Sabah Ansar, Mary L. Michaelis, Jennifer L. Bean, Rick T DobrowskyAbstract:Ceramide is a bioactive sphingolipid that can prevent calpain activation and beta-amyloid (A beta) neurotoxicity in cortical neurons. Recent evidence supports A beta induction of a calpain-dependent cleavage of the cyclin-dependent kinase 5 (cdk5) regulatory Protein P35 that contributes to tau hyperphosphorylation and neuronal death. Using cortical neurons isolated from wild-type and P35 knockout mice, we investigated whether ceramide required P35/cdk5 to protect against A beta-induced cell death and tau phosphorylation. Ceramide inhibited A beta-induced calpain activation and cdk5 activity in wild-type neurons and protected against neuronal death and tau hyperphosphorylation. Interestingly, A beta also increased cdk5 activity in P35-/- neurons, suggesting that the alternate cdk5 regulatory Protein, p39, might mediate this effect. In P35 null neurons, ceramide blocked A beta-induced calpain activation but did not inhibit cdk5 activity or cell death. However, ceramide blocked tau hyperphosphorylation potentially via inhibition of glycogen synthase kinase-3beta. These data suggest that ceramide can regulate A beta cell toxicity in a P35/cdk5-dependent manner.
Jigang Chen - One of the best experts on this subject based on the ideXlab platform.
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Identification and characterization of host cell Proteins interacting with Scylla serrata reovirus non-structural Protein P35
Virus Genes, 2016Co-Authors: Yangyang Yuan, Dongyang Fan, Sidong Zhu, Jifang Yang, Jigang ChenAbstract:We have previously shown that non-structural Protein P35, encoded by Scylla serrata reovirus (SsRV) S10, may act as a viroporin. To characterize the role of P35 Protein in the modulation of cellular function, a yeast two-hybrid system was used to screen a cDNA library derived from S. serrata to find its interacting partner. Protein interactions were confirmed in vitro by GST pull-down. Full cDNAs of P35 interactors were cloned by the rapid amplification of cDNA ends. After two-hybrid library screening, we isolated partial cDNAs encoding hemocyanin, cryptocyanin, and TAX1BP1. Interaction of P35 with each of hemocyanin, cryptocyanin, and TAX1BP1 was confirmed by GST pull-down. The full-length cDNA fragments of hemocyanin, cryptocyanin, and TAX1BP1 were 2287, 2422, and 3437 bp, respectively, and they encoded three putative Proteins with molecular masses of ~76.9, ~79.2, and ~107.2 kDa, respectively. This study casts new light on the function and physiological significance of P35 during the SsRV replication cycle.
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Scylla serrata reovirus P35 Protein expressed in Escherichia coli cells alters membrane permeability.
Virus Genes, 2015Co-Authors: Zhao Zhang, Yangyang Yuan, Dongyang Fan, Jifang Yang, Zhijuan Mao, Yan Yan, Jigang ChenAbstract:To promote viral entry, replication, release, and spread to neighboring cells, many cytolytic animal viruses encode Proteins responsible for modification of host cell membrane permeability and for formation of ion channels in host cell membranes. Scylla serrata reovirus (SsRV) is a major pathogen that can severely damage mud crab (S. serrata) aquaculture. Protein P35, which is encoded by segment 10 of SsRV, contains two transmembrane domains. In this study, we found that SsRV P35 can induce membrane permeability changes when expressed in Escherichia coli. SsRV P35 expressed in bacterial cells existed as monomers under reducing conditions but formed homodimers and homotrimers under non-reducing conditions. These findings demonstrate that P35 may act as a viroporin; further studies are needed to elucidate the detailed structure–function relationships of this Protein.
David J. Davido - One of the best experts on this subject based on the ideXlab platform.
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Herpes Simplex Virus 1 Upregulates P35, Alters CDK-5 Localization, and Stimulates CDK-5 Kinase Activity during Acute Infection in Neurons
Journal of Virology, 2015Co-Authors: Heba H. Mostafa, Jessica M. Van Loben Sels, David J. DavidoAbstract:The cyclin-dependent kinase 5 (CDK-5) activating Protein, P35, is important for acute herpes simplex virus 1 (HSV-1) replication in mice. This report shows that HSV-1 increases P35 levels, changes the primary localization of CDK-5 from the nucleus to the cytoplasm, and enhances CDK-5 activity during lytic or acute infection. Infected neurons also stained positive for the DNA damage response (DDR) marker γH2AX. We propose that CDK-5 is activated by the DDR to protect infected neurons from apoptosis.
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Role of a cdk5-associated Protein, P35, in herpes simplex virus type 1 replicationin vivo
Journal of NeuroVirology, 2010Co-Authors: Steve D Haenchen, Jeff A Utter, Adam M Bayless, Rick T Dobrowsky, David J. DavidoAbstract:Previous studies have shown that herpes simplex virus type 1 (HSV-1) replication is inhibited by the cyclin-dependent kinase (cdk) inhibitor roscovitine. One roscovitine-sensitive cdk that functions in neurons is cdk5, which is activated in part by its binding partner, P35. Because HSV establishes latent infections in sensory neurons, we sought to determine the role P35 plays in HSV-1 replication in vivo . For these studies, wild-type (wt) and P35-/- mice were infected with HSV-1 using the mouse ocular model of HSV latency and reactivation. The current results indicate that P35 is an important determinant of viral replication in vivo .
