Proliferation Defects and Genome Instability in Cells Lacking Cul4A

The Cul4A gene, which encodes a core component of a cullin-based E3 ubiquitin ligase complex, is over-expressed in breast and hepatocellular cancers. In breast cancers, over-expression of Cul4A strongly correlates with poor prognosis. Also, Cul4A is required for early embryonic development. Early lethality of mouse embryos prevented a detailed analysis of the functions of Cul4A. Here, we used a strain of mice carrying floxed alleles of Cul4A to study its role in cell division, in vitro and in vivo. Embryonic fibroblasts exhibit a severe deficiency in cell proliferation following deletion of Cul4A. We observed that the Cul4A protein is abundantly expressed in brain, liver and in the mammary tissue of pregnant mice. Deletion of Cul4A in liver impairs hepatocyte proliferation during regeneration following carbon tetrachloride induced injury. The Cul4A-deleted cells are slow in entering S phase, and are deficient in progressing through early M phase. Several cell cycle regulators, including p53 and p27Kip1, are de-regulated in the Cul4A-deleted cells. Expression of a dominant negative mutant of p53 causes significant reversal of the proliferation defects in Cul4A-deleted cells. The Cul4A-deleted cells exhibit aberrant number of centrosome, multipolar spindles and micronuclei formation. Furthermore, those cells are sensitive to UV irradiation and exhibit reduced levels of unscheduled DNA synthesis. Together, our observations indicate that Cul4A is required for efficient cell proliferation, control of the centrosome amplification and genome stability.