Degradation of the interferon-induced 68,000-Mr protein kinase by poliovirus requires RNA

Tracy L. Black, Glen N Barber, Michael G. Katze

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Abstract

Control of the interferon-induced double-stranded RNA (dsRNA) activated protein kinase (referred to as P68 because of its Mr of 68,000 in human cells) by animal viruses is essential to avoid decreases in protein synthetic rates during infection. We have previously demonstrated that poliovirus establishes a unique way of regulating the protein kinase, namely by inducing the specific degradation of P68 during infection (T. L. Black, B. Safer, A. Hovanessian, and M. G. Katze, J. Virol. 63:2244-2251, 1989). In the present study we investigated the mechanisms by which P68 degradation occurred. To do this we used an in vitro degradation assay which faithfully reproduced the in vivo events. Although viral gene expression was required for P68 degradation, the major poliovirus proteases, 2A and 3C, were found not to be directly involved with P68 proteolysis. However, the protease responsible for P68 degradation required divalent cations for maximal activity and probably has both an RNA and a protein component since trypsin and ribonuclease abrogated the activity. Despite this requirement for divalent cations and RNA, activation of the kinase was not required for proteolysis since a catalytically inactive P68 was still degraded. Mapping of P68 protease-sensitive sites by using in vitro translated truncation and deletion mutants revealed that sites required for degradation resided in the amino terminus and colocalized to dsRNA-binding domains. Finally, we found that preincubation of cell extracts with the synthetic dsRNA poly(I-C) largely prevented P68 proteolysis, providing additional evidence for the critical role of RNA. On the basis of these data, we present a hypothetical model depicting possible mechanisms of P68 degradation in poliovirus-infected cells.

Original languageEnglish
Pages (from-to)791-800
Number of pages10
JournalJournal of Virology
Volume67
Issue number2
StatePublished - Feb 1 1993
Externally publishedYes

Fingerprint

Enterovirus C
Poliovirus
interferons
protein kinases
Protein Kinases
Interferons
Proteolysis
Double-Stranded RNA
Divalent Cations
RNA
degradation
Peptide Hydrolases
double-stranded RNA
eIF-2 Kinase
proteolysis
Viral Genes
Ribonucleases
Infection
proteinases
Cell Extracts

ASJC Scopus subject areas

  • Immunology

Cite this

Degradation of the interferon-induced 68,000-Mr protein kinase by poliovirus requires RNA. / Black, Tracy L.; Barber, Glen N; Katze, Michael G.

In: Journal of Virology, Vol. 67, No. 2, 01.02.1993, p. 791-800.

Research output: Contribution to journalArticle

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