Molecular cloning and characterization of RBCK2, a splicing variant of a RBCC family protein, RBCK1

RBCK1 (RBCC protein interacting with PKC 1) has two coiled-coil regions, a RING finger, a B-box and a B-box-like motif. RBCK2, a cDNA fragment related to RBCK1 was obtained, that lacks the 161-bp sequence of RBCK1 and encodes 260 amino acid residues. The 240-amino acid sequence in the NH2-terminal of RBCK2 is identical with RBCK1 and contains two coiled-coil regions but no other structural motifs, whereas the 20-amino acid sequence in the COOH-terminal is distinct from RBCK1. The analysis of genomic DNA revealed that RBCK1 and RBCK2 are generated from a single gene by alternative splicing. The RBCK1 protein interacted with the RBCK1 and RBCK2 proteins, but the RBCK2 protein did not interact with itself, in vitro. The RBCK2 protein fused with the DNA-binding domain of yeast GAL4 (GAL4DBD) did not show a transcriptional activity, but the RBCK2 protein inhibited the transcriptional activity of the RBCK1 protein fused with GAL4DBD. These results suggest that RBCK2 may inhibit the transcriptional activity of RBCK1 probably through complex formation with RBCK1.