The Role of Viral and Host Cell Proteins in Paramyxovirus Transcription and Replication

Moyer, Sue A.; Horikami, Sandra M.

doi:10.1007/978-1-4615-3790-8_9

Cited by 16 publications

(15 citation statements)

References 108 publications

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“…By contrast, although addition of purified tubulin alone to SeV particles led to the synthesis of leader-like RNA and mRNA for the 5Ј-proximal NP gene, downstream mRNAs were not transcribed. Similar results were reported for measles virus (12,16). Thus it seems that an additional factor(s) may be required for complete transcription of these viral genomes.…”

Section: Discussionsupporting

confidence: 77%

“…54). Previous studies suggested that the carboxyl-terminal tails of the tubulin subunits, which are highly negatively charged and exposed on the surface of the tubules, may be involved in SeV transcription (16,18). It is important to investigate the significance of the acidic tails of tubulin in the transcription-stimulatory activity and the recruitment of other host factors such as PGK.…”

Section: Discussionmentioning

confidence: 99%

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Involvement of a Cellular Glycolytic Enzyme, Phosphoglycerate Kinase, in Sendai Virus Transcription

Ogino¹,

Iwama²,

Kinouchi³

et al. 1999

Journal of Biological Chemistry

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In vitro mRNA synthesis of Sendai virus is almost entirely dependent on the addition of cellular proteins (host factors). Previous studies indicated that the host factor activity from bovine brain was resolved into at least two complementary fractions, one of which may be tubulin. In this study, the host factor activity that stimulates the transcription in the presence of tubulin was further purified from bovine brain. This fraction was found to contain at least two complementary factors, and one of them was purified to a single polypeptide chain with an apparent M r of 46,000 (p46). From the amino acid sequence, biochemical, and immunological analyses, p46 was identified as a glycolytic enzyme, phosphoglycerate kinase (PGK). Purified native PGK from rabbit and yeast, and a recombinant human PGK substituted for p46. Although, as previously suggested, tubulin was involved in the transcription initiation complex formation by being integrated into the complex, p46 and its complementary factor had little effect on the complex formation. On the other hand, when p46 and the complementary factor were added to the RNA chain elongation reaction from the isolated initiation complex formed with tubulin, mRNA synthesis was dramatically stimulated. The enzymatic activity per se of PGK did not seem to be required for its activity. West-Western blot analysis showed that PGK could directly interact with tubulin. These data suggest that PGK stimulates Sendai virus mRNA synthesis at the elongation step, probably through its interaction with tubulin in the initiation complex.

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Section: Discussionsupporting

confidence: 77%

Section: Discussionmentioning

confidence: 99%

Involvement of a Cellular Glycolytic Enzyme, Phosphoglycerate Kinase, in Sendai Virus Transcription

Ogino¹,

Iwama²,

Kinouchi³

et al. 1999

Journal of Biological Chemistry

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“…The DI-H genome RNA is 1,411 nucleotides long (about 9% the size of the nondefective genome), contains sequences from the 5' end of the genome RNA, including a portion of the L cistron, and has copy-back termini, with 110 nucleotides of the 3' terminus complementary to the 5' terminus (2,19). In previous work we showed that purified NP protein would support the replication of intracellular DI-H nucleocapsids, but not of purified virus, which required the addition of infected cell extracts for nucleocapsid RNA replication (1,22). These data suggested that NP protein alone was sufficient for the elongation and encapsidation of preinitiated replicating RNA present in intracellular nucleocapsids but that additional viral proteins * Corresponding author.…”

mentioning

confidence: 98%

“…We have used the Sendai virus defective interfering particle DI-H as the template for the study of genome replication (1,3,22). The DI-H genome RNA is 1,411 nucleotides long (about 9% the size of the nondefective genome), contains sequences from the 5' end of the genome RNA, including a portion of the L cistron, and has copy-back termini, with 110 nucleotides of the 3' terminus complementary to the 5' terminus (2,19).…”

mentioning

confidence: 99%

Complexes of Sendai virus NP-P and P-L proteins are required for defective interfering particle genome replication in vitro

Horikami

Curran

Kolakofsky

et al. 1992

J Virol

230

108

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We present evidence that the formation of NP-P and P-L protein complexes is essential for replication of the genome of Sendai defective interfering (DI-H) virus in vitro, using extracts of cells expressing these viral proteins from plasmids. Optimal replication of DI-H nucleocapsid RNA required extracts of cells transfected with critical amounts and ratios of each of the plasmids and was threeto fivefold better than replication with a control extract prepared from a natural virus infection. Extracts in which NP and P proteins were coexpressed supported replication of the genome of purified DI-H virus which contained endogenous polymerase proteins, but extracts in which NP and P were expressed separately and then mixed were inactive. Similarly, the P and L proteins must be coexpressed for biological activity. The replication data thus suggest that two protein complexes, NP-P and P-L, are required for nucleocapsid RNA replication and that these complexes must form during or soon after synthesis of the proteins. Biochemical evidence in support of the formation of each complex includes coimmunoprecipitation of both proteins of each complex with an antibody specific for one component and cosedimentation of the subunits of each complex. We propose that the P-L complex serves as the RNA polymerase and NP-P is required for encapsidation of newly synthesized RNA.

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“…For VSV, a leader-to-N-transcript molar ratio of ϳ7.0 was obtained by using transcription reconstitution conditions with optimal amounts of L and P proteins (8) and a ratio of ϳ2.9 was obtained by using detergent-disrupted virions (11). For Sendai virus, roughly equimolar synthesis was found in one case (37) and a large excess of leader over N transcript (ratio, Ն64) was found in the other (23). The wide range in measured ratios is perhaps not surprising in light of several studies showing preferential inhibition of mRNA synthesis relative to leader synthesis under a variety of reaction conditions, including low ATP concentration (2), replacement of ATP with the nonhydrolyzable analog ␤,␥-imido ATP (13,26), suboptimal amounts of P protein (8), and addition of a monoclonal antibody reactive with both P and N proteins (39).…”

mentioning

confidence: 99%

Initiation of vesicular stomatitis virus mutant polR1 transcription internally at the N gene in vitro

Chuang

Perrault

1997

J Virol

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The vesicular stomatitis virus (VSV) polymerase is thought to initiate transcription of its genome by first copying a small leader RNA complementary to the 3 end of the template. The polR VSV mutants, in contrast to wild-type virus, frequently read through the leader termination site during transcription in vitro. To shed light on polymerase termination and reinitiation events at the crucial leader-N gene junction, we employed RNase protection assays to precisely measure molar ratios of leader, N, and readthrough transcript accumulation in vitro. Wild-type virus synthesized essentially equimolar amounts of leader and N transcripts, but, unexpectedly, the polR1 mutant yielded about twice as much N mRNA as leader (ratio of 1.9 ؎ 0.1). Primer extension assays ruled out an increase in abortive N transcript synthesis for polR1. Transcription entailed multiple rounds of synthesis, with transcript ratios remaining the same after 0.5 or 2 h of synthesis, ruling out a significant contribution from polymerases "pre-positioned" at the N gene. No significant degradation of either leader or N transcripts was observed after incubating purified products with virions. Our data lead us to conclude that transcription can initiate internally at the N gene, at least in the case of polR1 VSV. We propose, however, that productive internal initiation of transcription is a fundamental property of the VSV polymerase and that of related viruses. A model postulating two distinct polymerase complexes, one for leader synthesis and one for internal initiation, is presented.

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The Role of Viral and Host Cell Proteins in Paramyxovirus Transcription and Replication

Cited by 16 publications

References 108 publications

Involvement of a Cellular Glycolytic Enzyme, Phosphoglycerate Kinase, in Sendai Virus Transcription

Involvement of a Cellular Glycolytic Enzyme, Phosphoglycerate Kinase, in Sendai Virus Transcription

Complexes of Sendai virus NP-P and P-L proteins are required for defective interfering particle genome replication in vitro

Initiation of vesicular stomatitis virus mutant polR1 transcription internally at the N gene in vitro

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