DOI . ORG {
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We began analyzing https://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1000054, but it redirected us to https://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1000054. The analysis below is for the second page.
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Author SummaryThe SARS-coronavirus (SARS-CoV), which causes the life-threatening severe acute respiratory syndrome, replicates in the cytoplasm of infected host cells. A critical early step in the SARS-CoV life cycle is the formation of a replication/transcription complex (RTC) that drives viral genome replication and subgenomic mRNA synthesis. Virus-encoded enzymes form the core of this RTC, which is believed to be associated with characteristic virus-induced membrane structures derived from modified host cell membranes. To investigate the connection between these membrane structures and SARS-CoV RNA synthesis, and to characterize RTC composition and function, we isolated these complexes and developed the first in vitro assay to study their activity. SARS-CoV genomic RNA and all eight subgenomic mRNAs were synthesized in this in vitro reaction. By centrifugation, RTC activity could be isolated from the cytoplasm, together with membrane structures, viral enzymes, and RNA. The activity of these isolated RTCs was dependent on a cytoplasmic host factor. RTC activity was destroyed by detergent treatment, suggesting a critical role for membranes that appeared to protect the complex against protease and nuclease digestion. Our data establish a functional connection between viral RNA synthesis and intracellular membranes and show that host factors play a crucial role in SARS-CoV RNA synthesis.
Title[redir]:
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Description:
Author SummaryThe SARS-coronavirus (SARS-CoV), which causes the life-threatening severe acute respiratory syndrome, replicates in the cytoplasm of infected host cells. A critical early step in the SARS-CoV life cycle is the formation of a replication/transcription complex (RTC) that drives viral genome replication and subgenomic mRNA synthesis. Virus-encoded enzymes form the core of this RTC, which is believed to be associated with characteristic virus-induced membrane structures derived from modified host cell membranes. To investigate the connection between these membrane structures and SARS-CoV RNA synthesis, and to characterize RTC composition and function, we isolated these complexes and developed the first in vitro assay to study their activity. SARS-CoV genomic RNA and all eight subgenomic mRNAs were synthesized in this in vitro reaction. By centrifugation, RTC activity could be isolated from the cytoplasm, together with membrane structures, viral enzymes, and RNA. The activity of these isolated RTCs was dependent on a cytoplasmic host factor. RTC activity was destroyed by detergent treatment, suggesting a critical role for membranes that appeared to protect the complex against protease and nuclease digestion. Our data establish a functional connection between viral RNA synthesis and intracellular membranes and show that host factors play a crucial role in SARS-CoV RNA synthesis.
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Doi.org employs APACHE LENYA.
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Keywords {๐}
rna, nsp, rtc, sarscov, activity, fig, article, view, google, scholar, membrane, cells, fraction, synthesis, viral, vitro, structures, coronavirus, host, virus, pns, membranes, replication, rnas, protein, lane, synthesized, ivra, virol, cytoplasmic, min, analyzed, rtcs, reaction, proteins, plos, replicase, presence, lanes, factor, protease, genome, newly, nsps, sarscovinfected, data, image, treatment, severe, acute,
Topics {โ๏ธ}
molecular biology plos pathog 4 cookies plos classical genetics euro-asian sars-dtv network open-access article distributed sars-cov-induced membrane structures sars-coronavirus replication/transcription complexes sars-cov life cycle sars-cov rna synthesis sars-cov-induced nsp cross-reacting host protein host protein cross-reacting murine coronavirus-infected cells virus-induced membrane structures viral rna-synthesizing machinery virus-induced structures derived cytoplasmic nucleases double-stranded rna intermediates sars-cov genomic rna cellular nucleases 32p-labeled oligonucleotide sarsv002 sars-cov-infected cells sars-cov-infected cells [8] sars-cov infected cells active sars-cov rtcs 35s-labeled amino acids protein synthesis occurred coronavirus rna synthesis sars-cov titer present 32p-labeled rna molecules negative strand-specific probe 32p-labeled rna products rtc-related membrane rearrangements coronavirus protein processing active sars-cov rtc original author viral life cycle positive strand-specific probe viral rna synthesis virus-encoded enzymes form sars-coronavirus genome sars-cov rtc activity nucleases suggesting product-specific differences rna synthesis occurs membrane-protected replication complexes replication/transcription complex ๏ฟฝreplication/transcription complex 10 ยตci [ฮฑ-32p]ctp
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