Cell cycle regulation of hepatitis C and encephalomyocarditis virus internal ribosome entry site-mediated translation in human embryonic kidney 293 cells

Arun Venkatesan, Rakhi Sharma, Asim Dasgupta

Research output: Contribution to journalArticlepeer-review

Abstract

We have established stably transformed human embryonic kidney cell lines (HEK293) containing bicistronic constructs to study regulation of viral internal ribosome entry site (IRES)-mediated translation in vivo. These cells produce Renilla luciferase (Rluc) in a cap-dependent manner, while Firefly luciferase (Luc) synthesis is mediated by IRES elements. Using these cell lines, we demonstrate here that IRES-mediated translation directed by both hepatitis C (HCV) and encephalomyocarditis (EMCV) virus varies with the cell cycle. Experiments involving arrest of the cell lines at different phases of the cell cycle, release of synchronized cells from cell cycle arrest, as well as direct sorting of the cells based on position in the cell cycle have shown that the activity of the HCV and EMCV IRES elements is lowest during the G2/M phase in HEK293 cells. These results suggest that cellular trans-acting factors either stimulate viral IRES-mediated translation during G1 and S phases or repress translation during the G2/M phase in HEK293 cells.

Original languageEnglish (US)
Pages (from-to)85-95
Number of pages11
JournalVirus Research
Volume94
Issue number2
DOIs
StatePublished - Aug 1 2003
Externally publishedYes

Keywords

  • Cell cycle
  • Encephalomyocarditis virus
  • HEK 293 cells
  • Hepatitis C

ASJC Scopus subject areas

  • Cancer Research
  • Virology
  • Infectious Diseases

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