Sequences of the vesicular stomatitis virus matrix protein involved in binding to nucleocapsids

Kaptur, Paulina E.; Rhodes, Richard B.; Lyles, Douglas S.

doi:10.1128/jvi.65.3.1057-1065.1991

Cited by 40 publications

(34 citation statements)

References 25 publications

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“…We used limited proteolysis of M protein with trypsin or S. aureus V8 protease to determine which M protein fragments contained phosphate residues. Proteolytic cleavage sites in VSV M protein were previously determined by using aminoterminal sequencing by Edman degradation (14,27). Table 1 shows the sequences for the 50 N-terminal amino acids for ts23rl M protein, a temperature-stable revertant of the ts M protein mutant tsO23 (24), and for the San Juan strain M protein (28).…”

Section: Resultsmentioning

confidence: 99%

“…Purification of nucleocapsid-M protein complexes. Nucleocapsid-M protein complexes were purified as described previously (14). Briefly, purified virus was solubilized with Triton X-100 in low-ionic-strength buffer and then fractionated on a discontinuous sucrose gradient in the absence of detergent.…”

Section: Materuils and Methodsmentioning

confidence: 99%

“…Phosphoamino acid analysis. Either purified 32P-labeled virions or purified 32P-labeled nucleocapsid-M protein complexes were subjected to PAGE, then electroblotted onto polyvinylidene difluoride (PVDF) membranes (13), and stained with Coomassie blue as described previously (14). M and NS protein bands were excised and subjected to vaporphase acid hydrolysis.…”

Section: Materuils and Methodsmentioning

confidence: 99%

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Sites of in vivo phosphorylation of vesicular stomatitis virus matrix protein

Kaptur

McCreedy

Lyles

1992

J Virol

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We mapped the in vivo phosphorylation sites for the matrix (M) protein of the Orsay and San Juan strains of vesicular stomatitis virus, Indiana serotype, using limited proteolysis and phosphoamino acid analysis. M protein was solubilized from 32P-labeled virions by using detergent and high-salt conditions, then treated with either trypsin or Staphylococcus aureus V8 protease, and analyzed by polyacrylamide gel electrophoresis and autoradiography to determine which fragments contained phosphate residues. The M protein fragment extending from amino acid 20 to the carboxy terminus contained approximately 70% of the control 32p label, while the fragment extending from amino acid 35 to the carboxy terminus had only trace amounts of label. These data indicate that the major phosphorylation site was between amino acids 20 and 34 in the Orsay strain M protein. Phosphoamino acid analysis of M protein by thin-layer electrophoresis showed the presence of phosphothreonine and phosphoserine and that phosphothreonine continued to be released after prolonged vapor-phase acid hydrolysis. These data identify Thr-31 as the primary in vivo phosphate acceptor for M protein of the Orsay strain of vesicular stomatitis virus. The San Juan strain M protein has serine at position 32, which may also be an important phosphate acceptor. In addition, phosphorylation at Ser-2,-3, or-17 occurs to a greater extent in the San Juan strain M protein than in the Orsay strain M protein. The subcellular distribution of phosphorylated M protein was investigated to determine a probable intracellular site(s) of phosphorylation. Phosphorylated M protein was associated primarily with cellular membranes, suggesting phosphorylation by a membrane-associated kinase. Virion M protein was phosphorylated to a greater extent than membrane-bound M protein, indicating that M protein phosphorylation occurs at a late stage in virus assembly. Phosphorylation of wild-type and temperature-sensitive mutant M protein was studied in vivo at the nonpermissive temperature. The data show that phosphorylated M protein was detected only in wild-type virus-infected cells and virions, suggesting that association with nucleocapsids may be required for M protein phosphorylation or that misfolding of mutant M protein at the nonpermissive temperature prevents

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Section: Resultsmentioning

confidence: 99%

Section: Materuils and Methodsmentioning

confidence: 99%

Section: Materuils and Methodsmentioning

confidence: 99%

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Sites of in vivo phosphorylation of vesicular stomatitis virus matrix protein

Kaptur

McCreedy

Lyles

1992

J Virol

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“…Another cis-acting viral element(s) also may be involved in the packaging of nucleocapsid (81). Rhabdovirus matrix protein interacts with the viral helical nucleocapsid (38,61), and this interaction probably is important for the packaging of helical nucleocapsid into virus particles, although the mechanism of the selective recognition of nucleocapsid containing the genomic RNA by the matrix protein is unknown. For the negative-strand RNA viruses carrying the genome in a helical nucleocapsid, association of nucleocapsid protein with RNA appears to be a prerequisite for RNA packaging, but a mechanism for selective packaging of specific intracellular helical nucleocapsids is not described.…”

Section: Discussionmentioning

confidence: 99%

Cooperation of an RNA Packaging Signal and a Viral Envelope Protein in Coronavirus RNA Packaging

Narayanan

Makino

2001

J Virol

110

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Murine coronavirus mouse hepatitis virus (MHV) produces a genome-length mRNA, mRNA 1, and six or seven species of subgenomic mRNAs in infected cells. Among these mRNAs, only mRNA 1 is efficiently packaged into MHV particles. MHV N protein binds to all MHV mRNAs, whereas envelope M protein interacts only with mRNA 1. This M protein-mRNA 1 interaction most probably determines the selective packaging of mRNA 1 into MHV particles. A short cis-acting MHV RNA packaging signal is necessary and sufficient for packaging RNA into MHV particles. The present study tested the possibility that the selective M protein-mRNA 1 interaction is due to the packaging signal in mRNA 1. Regardless of the presence or absence of the packaging signal, N protein bound to MHV defective interfering RNAs and intracellularly expressed non-MHV RNA transcripts to form ribonucleoprotein complexes; M protein, however, interacted selectively with RNAs containing the packaging signal. Moreover, only the RNA that interacted selectively with M protein was efficiently packaged into MHV particles. Thus, it was the packaging signal that mediated the selective interaction between M protein and viral RNA to drive the specific packaging of RNA into virus particles. This is the first example for any RNA virus in which a viral envelope protein and a known viral RNA packaging signal have been shown to determine the specificity and selectivity of RNA packaging into virions.Within the "soup" of a virally infected cell, viral genome and viral proteins specifically and selectively coalesce into progeny viruses. The process of packaging the viral genome, or surrounding the nucleic acid with protein and possibly an envelope, is a critical step in production of new virus. Within their hosts, RNA viruses manufacture genomic RNA, antigenomic RNA, and, in some cases, subgenomic-length RNAs all in the presence of ubiquitous host cell mRNAs, tRNAs, and rRNAs. Occasional packaging of nongenomic viral RNAs and cellular RNAs results in noninfectious viruses, and yet this packaging seems to occur at constant rates which are characteristic for different species of viruses. Unchecked packaging of cellular nucleic acid into viral particles would be expected to overwhelm the ability of intracellular viral genomic RNA to associate with limited viral and host assembly factors. Each virus, therefore, probably has developed a defensive strategy for specific and selective packaging of intracellular genomic RNA into virus particles. RNA packaging signals required for viral RNA packaging are known for several RNA viruses (1,3,6,9,26,28,48,65,66,81), and for some of these, the packaging signal is all that is needed (1,66,80,82).A critical step for the selective packaging of viral genomic RNA in those RNA viruses with icosahedral and spherical cores is the binding of core protein to intracellular genomic RNA; only the viral RNAs that associate with core protein are packaged into virus particles. The case for negative-strand RNA viruses with a helical nucleocapsid structure seems to be tha...

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“…The M proteins of mononegaviruses are thought to have two key roles: to inhibit the transcriptase activity of the nucleocapsid prior to packaging, and to mediate association of the nucleocapsid with the nascent viral envelope (Kaptur et al, 1991;Lenard, 1996;Coronel et al, 2001). The functions of RSV M in infected cells have not been examined in detail (Ghildyal et al, 2002;Henderson et al, 2002;Rodriguez et al, 2004), but by analogy with the M proteins of other paramyxoviruses as well as VSV, RSV M is presumed to play a crucial role in virus assembly, budding and the formation of virus particles.…”

Section: Proteinmentioning

confidence: 99%

Central role of the respiratory syncytial virus matrix protein in infection

Ghildyal

Jans

2006

FEMS Microbiol Rev

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Respiratory syncytial virus is the major respiratory pathogen of infants and children worldwide, with no effective treatment or vaccine available. Steady progress has been made in understanding the respiratory syncytial virus life cycle and the consequences of infection, but many areas of respiratory syncytial virus biology remain poorly understood, including the role of subcellular localisation of respiratory syncytial virus gene products such as the matrix protein in the infected host cell. The matrix protein plays a central role in viral assembly and, intriguingly, has been observed to traffic into and out of the nucleus at specific times during the respiratory syncytial virus infectious cycle. Further, the matrix protein has been shown to be able to inhibit transcription, which may be a key to respiratory syncytial virus pathogenesis. This review will focus on the role of the matrix protein in respiratory syncytial virus infection and what is known of its nucleocytoplasmic trafficking, the understanding of which may lead to new therapeutic approaches to combat respiratory syncytial virus, and/or vaccine development.

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Sequences of the vesicular stomatitis virus matrix protein involved in binding to nucleocapsids

Cited by 40 publications

References 25 publications

Sites of in vivo phosphorylation of vesicular stomatitis virus matrix protein

Sites of in vivo phosphorylation of vesicular stomatitis virus matrix protein

Cooperation of an RNA Packaging Signal and a Viral Envelope Protein in Coronavirus RNA Packaging

Central role of the respiratory syncytial virus matrix protein in infection

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