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Molecular and Cellular Biology, April 2003, p. 3013-3028, Vol. 23, No. 8
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.8.3013-3028.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

CDP/Cux Stimulates Transcription from the DNA Polymerase {alpha} Gene Promoter

Mary Truscott,1,2 Lélia Raynal,1 Peter Premdas,1 Brigitte Goulet,1,2 Lam Leduy,1 Ginette Bérubé,1 and Alain Nepveu1,2,3,4*

Molecular Oncology Group, McGill University Health Center,1 Departments of Biochemistry,2 Medicine,3 Oncology, McGill University, Montreal, Quebec, Canada H3A 1A14

Received 15 August 2002/ Returned for modification 30 September 2002/ Accepted 23 January 2003

CDP/Cux (CCAAT-displacement protein/cut homeobox) contains four DNA binding domains, namely, three Cut repeats (CR1, CR2, and CR3) and a Cut homeodomain. CCAAT-displacement activity involves rapid but transient interaction with DNA. More stable DNA binding activity is up-regulated at the G1/S transition and was previously shown to involve an N-terminally truncated isoform, CDP/Cux p110, that is generated by proteolytic processing. CDP/Cux has been previously characterized as a transcriptional repressor. However, here we show that expression of reporter plasmids containing promoter sequences from the human DNA polymerase {alpha} (pol {alpha}), CAD, and cyclin A genes is stimulated in cotransfections with N-terminally truncated CDP/Cux proteins but not with full-length CDP/Cux. Moreover, expression of the endogenous DNA pol {alpha} gene was stimulated following the infection of cells with a retrovirus expressing a truncated CDP/Cux protein. Chromatin immunoprecipitation (ChIP) assays revealed that CDP/Cux was associated with the DNA pol {alpha} gene promoter specifically in the S phase. Using linker scanning analyses, in vitro DNA binding, and ChIP assays, we established a correlation between binding of CDP/Cux to the DNA pol {alpha} promoter and the stimulation of gene expression. Although we cannot exclude the possibility that stimulation of gene expression by CDP/Cux involved the repression of a repressor, our data support the notion that CDP/Cux participates in transcriptional activation. Notwithstanding its mechanism of action, these results establish CDP/Cux as an important transcriptional regulator in the S phase.

* Corresponding author. Mailing address: Molecular Oncology Group, McGill University Health Center, McGill University, 687 Pine Ave. West, Montreal, Quebec, Canada H3A 1A1. Phone: (514) 842-1231, ext. 35842. Fax: (514) 843-1478. E-mail: alain.nepveu{at}mcgill.ca.

Molecular and Cellular Biology, April 2003, p. 3013-3028, Vol. 23, No. 8
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.8.3013-3028.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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