E2F Mediates Dihydrofolate Reductase Promoter Activation and Multiprotein Complex Formation in Human Cytomegalovirus Infection

Michael Wade, Timothy F. Kowalik, Maria Mudryj, Eng Shang Huang, Jane Clifford Azizkhan

Research output: Contribution to journalArticle

96 Citations (Scopus)

Abstract

The adenovirus immediate-early protein E1A activates the adenovirus E2 promoter and several cellular gene promoters through transcription factor E2F. The immediate-early proteins of human cytomegalovirus (HCMV) can complement an E1A-deficient adenovirus mutant and activate the adenovirus E2 promoter. HCMV also has been shown to activate the adenovirus E2 promoter. On the basis of these findings, we have investigated whether HCMV can activate the promoter of the cellular dihydrofolate reductase (DHFR) gene, which requires E2F binding for maximal promoter activity. We show that HCMV activates the DHFR promoter and that products of the HCMV major immediate-early gene region mediate the activation of the promoter specifically through the E2F site. We used gel mobility shift assays to search for potential molecular mechanisms for this activation and found an "infection-specific" multimeric complex that bound to the E2F sites in the DHFR and E2 promoters in extracts from HCMV-infected cells but not in extracts from uninfected cells. Several antibodies against HCMV immediate-early gene products had no effect on this infection-specific complex. Subsequently, the complex was found to contain E2F, cyclin A, p33cdk2, and p107 and to be similar to S-phase-specific complexes that recently have been identified in several cell types. A functional role for the binding of the cyclin A-p33cdk2 complex to cellular gene promoters has yet to be demonstrated; however, HCMV infection causes the induction of both cellular DNA replication and transcription of growth-related genes containing E2F sites in thier promoters. The findings described above therefore may relate to both of these effects of HCMV infection. We also provide evidence that some of the molecular events associated with adenovirus infection are different from those associated with HCMV infection.

Original languageEnglish (US)
Pages (from-to)4364-4374
Number of pages11
JournalMolecular and Cellular Biology
Volume12
Issue number10
StatePublished - Oct 1992
Externally publishedYes

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Multiprotein Complexes
Tetrahydrofolate Dehydrogenase
Cytomegalovirus Infections
Cytomegalovirus
Adenoviridae
Cyclin A
Immediate-Early Genes
Adenovirus Early Proteins
Genes
Immediate-Early Proteins
E2F Transcription Factors
Adenoviridae Infections
Electrophoretic Mobility Shift Assay
Infection
Cell Extracts
DNA Replication
S Phase
Gels

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

E2F Mediates Dihydrofolate Reductase Promoter Activation and Multiprotein Complex Formation in Human Cytomegalovirus Infection. / Wade, Michael; Kowalik, Timothy F.; Mudryj, Maria; Huang, Eng Shang; Azizkhan, Jane Clifford.

In: Molecular and Cellular Biology, Vol. 12, No. 10, 10.1992, p. 4364-4374.

Research output: Contribution to journalArticle

Wade, Michael ; Kowalik, Timothy F. ; Mudryj, Maria ; Huang, Eng Shang ; Azizkhan, Jane Clifford. / E2F Mediates Dihydrofolate Reductase Promoter Activation and Multiprotein Complex Formation in Human Cytomegalovirus Infection. In: Molecular and Cellular Biology. 1992 ; Vol. 12, No. 10. pp. 4364-4374.
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