Human cytomegalovirus infection induces cellular thymidylate synthase gene expression in quiescent fibroblasts

Giorgio Gribaudo, Ludovica Riera, Thomas L. Rudge, Patrizia Caposio, Lee F. Johnson, Santo Landolfo

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Productive infection of non-proliferating cells by cytomegalovirus (CMV) requires the coordinated stimulation of host biochemical pathways that prepare cells to synthesize DNA. Here we illustrate the ability of human CMV (HCMV) to stimulate cellular thymidylate synthase (TS) gene expression in quiescent human embryonic lung fibroblasts. TS mRNA and protein levels are nearly undetectable in quiescent cells, but are greatly increased following HCMV infection. Inhibition of TS activity was shown to impair HCMV DNA synthesis, demonstrating that TS upregulation is required for efficient HCMV replication in quiescent cells. The increase in TS gene expression was due to an increase in gene transcription, since the expression of a reporter gene driven by the human TS promoter was strongly induced by HCMV infection. Deletion analysis of the human TS promoter identified two positive elements that are important for this increased transcription. We have previously shown that murine CMV (MCMV) stimulates the mouse TS promoter by a mechanism that depends on the presence of an E2F element in the promoter region. However, deletion of the two potential E2F binding sites in the human TS promoter did not prevent the virus-induced increase in TS promoter activity. Our data suggest that HCMV activates human TS gene transcription by mechanisms that independent of E2F and different from those used by MCMV to stimulate the mouse TS promoter.

Original languageEnglish (US)
Pages (from-to)2983-2993
Number of pages11
JournalJournal of General Virology
Volume83
Issue number12
StatePublished - Dec 1 2002
Externally publishedYes

Fingerprint

Thymidylate Synthase
Cytomegalovirus Infections
Fibroblasts
Gene Expression
Cytomegalovirus
Infection
Muromegalovirus
DNA
Reporter Genes
Genetic Promoter Regions
Genes
Up-Regulation

ASJC Scopus subject areas

  • Virology
  • Immunology

Cite this

Gribaudo, G., Riera, L., Rudge, T. L., Caposio, P., Johnson, L. F., & Landolfo, S. (2002). Human cytomegalovirus infection induces cellular thymidylate synthase gene expression in quiescent fibroblasts. Journal of General Virology, 83(12), 2983-2993.

Human cytomegalovirus infection induces cellular thymidylate synthase gene expression in quiescent fibroblasts. / Gribaudo, Giorgio; Riera, Ludovica; Rudge, Thomas L.; Caposio, Patrizia; Johnson, Lee F.; Landolfo, Santo.

In: Journal of General Virology, Vol. 83, No. 12, 01.12.2002, p. 2983-2993.

Research output: Contribution to journalArticle

Gribaudo, G, Riera, L, Rudge, TL, Caposio, P, Johnson, LF & Landolfo, S 2002, 'Human cytomegalovirus infection induces cellular thymidylate synthase gene expression in quiescent fibroblasts', Journal of General Virology, vol. 83, no. 12, pp. 2983-2993.
Gribaudo, Giorgio ; Riera, Ludovica ; Rudge, Thomas L. ; Caposio, Patrizia ; Johnson, Lee F. ; Landolfo, Santo. / Human cytomegalovirus infection induces cellular thymidylate synthase gene expression in quiescent fibroblasts. In: Journal of General Virology. 2002 ; Vol. 83, No. 12. pp. 2983-2993.
@article{d6c19e99db834bd88c8217b981a33d82,
title = "Human cytomegalovirus infection induces cellular thymidylate synthase gene expression in quiescent fibroblasts",
abstract = "Productive infection of non-proliferating cells by cytomegalovirus (CMV) requires the coordinated stimulation of host biochemical pathways that prepare cells to synthesize DNA. Here we illustrate the ability of human CMV (HCMV) to stimulate cellular thymidylate synthase (TS) gene expression in quiescent human embryonic lung fibroblasts. TS mRNA and protein levels are nearly undetectable in quiescent cells, but are greatly increased following HCMV infection. Inhibition of TS activity was shown to impair HCMV DNA synthesis, demonstrating that TS upregulation is required for efficient HCMV replication in quiescent cells. The increase in TS gene expression was due to an increase in gene transcription, since the expression of a reporter gene driven by the human TS promoter was strongly induced by HCMV infection. Deletion analysis of the human TS promoter identified two positive elements that are important for this increased transcription. We have previously shown that murine CMV (MCMV) stimulates the mouse TS promoter by a mechanism that depends on the presence of an E2F element in the promoter region. However, deletion of the two potential E2F binding sites in the human TS promoter did not prevent the virus-induced increase in TS promoter activity. Our data suggest that HCMV activates human TS gene transcription by mechanisms that independent of E2F and different from those used by MCMV to stimulate the mouse TS promoter.",
author = "Giorgio Gribaudo and Ludovica Riera and Rudge, {Thomas L.} and Patrizia Caposio and Johnson, {Lee F.} and Santo Landolfo",
year = "2002",
month = "12",
day = "1",
language = "English (US)",
volume = "83",
pages = "2983--2993",
journal = "Journal of General Virology",
issn = "0022-1317",
publisher = "Society for General Microbiology",
number = "12",

}

TY - JOUR

T1 - Human cytomegalovirus infection induces cellular thymidylate synthase gene expression in quiescent fibroblasts

AU - Gribaudo, Giorgio

AU - Riera, Ludovica

AU - Rudge, Thomas L.

AU - Caposio, Patrizia

AU - Johnson, Lee F.

AU - Landolfo, Santo

PY - 2002/12/1

Y1 - 2002/12/1

N2 - Productive infection of non-proliferating cells by cytomegalovirus (CMV) requires the coordinated stimulation of host biochemical pathways that prepare cells to synthesize DNA. Here we illustrate the ability of human CMV (HCMV) to stimulate cellular thymidylate synthase (TS) gene expression in quiescent human embryonic lung fibroblasts. TS mRNA and protein levels are nearly undetectable in quiescent cells, but are greatly increased following HCMV infection. Inhibition of TS activity was shown to impair HCMV DNA synthesis, demonstrating that TS upregulation is required for efficient HCMV replication in quiescent cells. The increase in TS gene expression was due to an increase in gene transcription, since the expression of a reporter gene driven by the human TS promoter was strongly induced by HCMV infection. Deletion analysis of the human TS promoter identified two positive elements that are important for this increased transcription. We have previously shown that murine CMV (MCMV) stimulates the mouse TS promoter by a mechanism that depends on the presence of an E2F element in the promoter region. However, deletion of the two potential E2F binding sites in the human TS promoter did not prevent the virus-induced increase in TS promoter activity. Our data suggest that HCMV activates human TS gene transcription by mechanisms that independent of E2F and different from those used by MCMV to stimulate the mouse TS promoter.

AB - Productive infection of non-proliferating cells by cytomegalovirus (CMV) requires the coordinated stimulation of host biochemical pathways that prepare cells to synthesize DNA. Here we illustrate the ability of human CMV (HCMV) to stimulate cellular thymidylate synthase (TS) gene expression in quiescent human embryonic lung fibroblasts. TS mRNA and protein levels are nearly undetectable in quiescent cells, but are greatly increased following HCMV infection. Inhibition of TS activity was shown to impair HCMV DNA synthesis, demonstrating that TS upregulation is required for efficient HCMV replication in quiescent cells. The increase in TS gene expression was due to an increase in gene transcription, since the expression of a reporter gene driven by the human TS promoter was strongly induced by HCMV infection. Deletion analysis of the human TS promoter identified two positive elements that are important for this increased transcription. We have previously shown that murine CMV (MCMV) stimulates the mouse TS promoter by a mechanism that depends on the presence of an E2F element in the promoter region. However, deletion of the two potential E2F binding sites in the human TS promoter did not prevent the virus-induced increase in TS promoter activity. Our data suggest that HCMV activates human TS gene transcription by mechanisms that independent of E2F and different from those used by MCMV to stimulate the mouse TS promoter.

UR - http://www.scopus.com/inward/record.url?scp=0036936420&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036936420&partnerID=8YFLogxK

M3 - Article

C2 - 12466474

AN - SCOPUS:0036936420

VL - 83

SP - 2983

EP - 2993

JO - Journal of General Virology

JF - Journal of General Virology

SN - 0022-1317

IS - 12

ER -