Isolation of the Rb-related p130 through its interaction with CDK2 and cyclins

Abstract

A two-hybrid protein interaction screen was used to isolate cDNAs encoding human proteins that can interact with human CDK2 in yeast. A new member of the retinoblastoma susceptibility gene family, Rbr-2 (Rb-related), was obtained. The sequence of the Rbr-2 protein shares ~50% identity with p107 and homology to Rb within the pocket domain. Several lines of evidence indicate that Rbr-2 is the adenovirus E1A-associated p130. Like Rb and p107, p130(Rbr-2) can bind to viral oncoproteins, SV40 large T antigen, and adenovirus E1A through its pocket domain. Although p130(Rbr-2) does not bind to CDK2 in vitro, it can interact with cyclins, with a clear preference for D-type cyclins. Because both CDK2 and p130(Rbr-2) show affinity for cyclins, we suggest that p130(Rbr-2) and CDK2 interacted through a yeast-derived cyclin bridge in the two-hybrid screen. The gene encoding p130(Rbr-2) mapped to 16q13, a region of frequent genomic alteration in human tumors.