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PMID: 1313918 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Infection of HeLa cells with poliovirus results in modification of a complex that binds to the rRNA promoter.

Journal of virology ·Vol. 66 ·No. 5 ·1992-05-00 ·Pages 3062-8

Rubinstein SJ, Hammerle T, Wimmer E, Dasgupta A

Abstract

In HeLa cells, RNA polymerase I (Pol I)-mediated transcription is severely inhibited soon after infection with poliovirus. We have developed a gel retardation assay to analyze DNA-protein complexes formed at the Pol I promoter. We show here that two complexes (A and C) formed by nuclear extracts from uninfected cells disappear after infection of cells with poliovirus. In contrast, a new, rapidly migrating complex (D) is formed in virus-infected cell extract. This change in the mobility of gel-retarded complexes correlates well with the kinetics of inhibition of rRNA transcription in virus-infected cells. Incubation of nuclear extracts from mock-infected cells with bacterially expressed, purified poliovirus protease 3C results in the disappearance of complexes A and C with concomitant generation of complex D. A partially purified transcription factor fraction derived from uninfected cells that contains complex A is able to restore Pol I transcription when added to virus-infected cell extracts, suggesting that this complex plays an important role in Pol I transcription. These results suggest that poliovirus proteinase 3C may have an important role in the shutoff of Pol I transcription in cells infected with poliovirus.

MeSH Terms
3C Viral Proteases Base Sequence Cell Nucleus/metabolism Cycloheximide/pharmacology Cysteine Endopeptidases/metabolism DNA-Binding Proteins/metabolism HeLa Cells Host-Parasite Interactions Humans Molecular Sequence Data Poliomyelitis/genetics Poliovirus/genetics Promoter Regions, Genetic/genetics RNA Polymerase I/biosynthesis RNA, Ribosomal/genetics Subcellular Fractions/metabolism Viral Proteins
Chemicals
DNA-Binding Proteins RNA, Ribosomal Viral Proteins Cycloheximide RNA Polymerase I Cysteine Endopeptidases 3C Viral Proteases 3C proteases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Rubinstein S J
Department of Microbiology and Immunology, UCLA School of Medicine.
Hammerle T
Wimmer E
Dasgupta A
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.66.5.3062-3068.1992
Published
1992-05-00
Pages
3062-8
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC241067
Subset
IM
Grants
NIAID NIH HHS · AI15122 · United States
NIAID NIH HHS · AI27451 · United States
NCI NIH HHS · CA28146 · United States
External Links
PubMed source DOI: 10.1128/JVI.66.5.3062-3068.1992 PMC Full Text View journal details
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