Repression of human cytomegalovirus major immediate early gene expression by the cellular transcription factor CCAAT displacement protein.
Initiation of human cytomegalovirus (HCMV) productive infection is dependent on the major immediate early (MIE) genes ie1 and ie2. Several putative binding sites for CCAAT displacement protein (CDP or CUX1) were identified within the MIE promoter/regulatory region. Binding assays demonstrated binding of CUX1 to MIE-region oligonucleotides containing the CUX1 core binding sequence ATCGAT and mutagenesis of this sequence abrogated CUX1 binding. Furthermore, CUX1 repressed expression of a luciferase reporter construct controlled by the MIE promoter, and mutation of CUX1 binding sites within the promoter diminished this repressive function of CUX1. In the context of virus infection of HEK293 cells transfected with the CUX1 expression vector, CUX1 showed evidence of association with the HCMV MIE regulatory region and inhibited the capacity of the virus to express ie1 and ie2 transcripts, suggesting that this cellular factor regulates MIE gene expression following virus entry. These data identify a role for CUX1 in repressing HCMV gene expression essential for initiation of the replicative cycle.
Stern
J Lewis
JL
Centre for Virus Research, Westmead Millennium Institute, PO Box 412, Westmead, New South Wales 2145, Australia.
Cao
John Z
JZ
Xu
Jiake
J
Mocarski
Edward S
ES
Slobedman
Barry
B
D019677:Artificial Gene Fusion; D002460:Cell Line; D003587:Cytomegalovirus / Q000502:physiology*; D015967:Gene Expression Regulation, Viral; D017781:Genes, Immediate-Early; D017930:Genes, Reporter; D018398:Homeodomain Proteins / Q000378:metabolism*; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D009687:Nuclear Proteins / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D012097:Repressor Proteins / Q000378:metabolism*; D014157:Transcription Factors
C080632:CUX1 protein, human; D018398:Homeodomain Proteins; D009687:Nuclear Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; D008156:Luciferases