PubAg

Main content area

Genome-wide location analysis and expression studies reveal a role for p110 CUX1 in the activation of DNA replication genes

Author:
Harada, Ryoko, Vadnais, Charles, Sansregret, Laurent, Leduy, Lam, Bérubé, Ginette, Robert, François, Nepveu, Alain
Source:
Nucleic acids research 2008 v.36 no.1 pp. 189-202
ISSN:
0305-1048
Subject:
transcription factors, DNA replication, retroviral vectors, microarray technology, antibodies, proteolysis, plasmids, gene expression, transcription (genetics), genes, small interfering RNA, chromatin, interphase, Human gammaherpesvirus 4
Abstract:
Proteolytic processing of the CUX1 transcription factor generates an isoform, p110 that accelerates entry into S phase. To identify targets of p110 CUX1 that are involved in cell cycle progression, we performed genome-wide location analysis using a promoter microarray. Since there are no antibodies that specifically recognize p110, but not the full-length protein, we expressed physiological levels of a p110 isoform with two tags and purified chromatin by tandem affinity purification (ChAP). Conventional ChIP performed on synchronized populations of cells confirmed that p110 CUX1 is recruited to the promoter of cell cycle-related targets preferentially during S phase. Multiple approaches including silencing RNA (siRNA), transient infection with retroviral vectors, constitutive expression and reporter assays demonstrated that most cell cycle targets are activated whereas a few are repressed or not affected by p110 CUX1. Functional classes that were over-represented among targets included DNA replication initiation. Consistent with this finding, constitutive expression of p110 CUX1 led to a premature and more robust induction of replication genes during cell cycle progression, and stimulated the long-term replication of a plasmid bearing the oriP replicator of Epstein Barr virus (EBV).
Agid:
2064982