Loss of a phosphorylated form of transcription factor CREB/ATF in poliovirus-infected cells

Steven Kliewer, Christian Muchardt, Richard Gaynor, Asim Dasgupta

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Host cell RNA synthesis is inhibited by poliovirus infection. We have studied the mechanism of poliovirusinduced inhibition of RNA polymerase II-mediated transcription by using the adenovirus early region 3 (E3) promoter. In vitro transcription from the E3 promoter was severely inhibited in extracts prepared from poliovirus-infected HeLa cells. Four regions in the E3 promoter have been shown to serve as binding sites for cellular transcription factors. These regions contain binding sites for transcription factors NF-1 (site IV), AP-1 (site III), CREB/ATF (site II), and the TATA factor (site I). Binding to these four regions was not significantly altered by poliovirus infection as assayed by DNase I footprinting analysis; furthermore, gel retardation assays failed to reveal dramatic differences in the total amount of CREB/ATF-, AP-1-, and NF-1-binding activity present in mock- or poliovirus-infected cell extracts. Gel retardation assays, however, did reveal significant qualitative differences in the DNA-protein complexes formed with a CREB/ATF-binding site in extracts prepared from poliovirus-infected cells as compared to mock-infected cell extracts. Radioimmunoprecipitation reactions performed with antiserum against CREB/ATF revealed a severe reduction in a phosphorylated form of the protein present in poliovirus-infected cell extracts. However, in vitro kinase reactions demonstrated that mock- and poliovirus-infected cell extracts contained similar levels of CREB/ATF. Expression from the E3 promoter was shown to be activated by CREB/ATF in vivo; this induction was dependent upon the phosphorylation of CREB/ATF. Thus, we propose that poliovirus infection inhibits transcription from the E3 promoter, at least in part, through the dephosphorylation of CREB/ATF.

Original languageEnglish (US)
Pages (from-to)4507-4515
Number of pages9
JournalJournal of Virology
Volume64
Issue number9
StatePublished - 1990

Fingerprint

Activating Transcription Factors
Enterovirus C
Poliovirus
transcription factors
Genetic Promoter Regions
Cell Extracts
promoter regions
extracts
cells
binding sites
transcription (genetics)
Binding Sites
Transcription Factor AP-1
Electrophoretic Mobility Shift Assay
Transcription Factors
Infection
gels
infection
deoxyribonuclease I
dephosphorylation

ASJC Scopus subject areas

  • Immunology

Cite this

Loss of a phosphorylated form of transcription factor CREB/ATF in poliovirus-infected cells. / Kliewer, Steven; Muchardt, Christian; Gaynor, Richard; Dasgupta, Asim.

In: Journal of Virology, Vol. 64, No. 9, 1990, p. 4507-4515.

Research output: Contribution to journalArticle

Kliewer, S, Muchardt, C, Gaynor, R & Dasgupta, A 1990, 'Loss of a phosphorylated form of transcription factor CREB/ATF in poliovirus-infected cells', Journal of Virology, vol. 64, no. 9, pp. 4507-4515.
Kliewer, Steven ; Muchardt, Christian ; Gaynor, Richard ; Dasgupta, Asim. / Loss of a phosphorylated form of transcription factor CREB/ATF in poliovirus-infected cells. In: Journal of Virology. 1990 ; Vol. 64, No. 9. pp. 4507-4515.
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