Interaction of rat Cdc37-related protein with retinoblastoma gene product.
By using in vitro binding assays and the yeast two-hybrid system, we have found that a full-length rat Cdc37-related protein (RCdc37) could associate specifically with the retinoblastoma susceptibility gene product (pRB). A series of GST-RCdc37 deletion mutants was constructed to define the amino acid sequence required for the interaction with pRB. A GST-RCdc37 (-20 approximately 229) possessed an activity to associate with pRB, whereas GST-RCdc37 (-20 approximately 165) lost its activity, indicating that the amino acid sequence between 166 and 229 of RCdc37 was essential for the association with pRB. Interestingly, there exists a highly conserved pRB-binding motif (LXCXE; X = any amino acid) that is essential for pRB binding of SV40 large T antigen, E1A, and E7 proteins. A similar experiment using a pRB deletion mutant revealed that the carboxy-terminal portion of pRB was required for binding to RCdc37.
Ozaki
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Division of Biochemistry, Chiba Cancer Center Research Institute, Japan.
Sakiyama
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D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002352:Carrier Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D017124:Conserved Sequence; D029721:Drosophila Proteins; D004926:Escherichia coli; D005982:Glutathione Transferase / Q000096:biosynthesis; D018832:Molecular Chaperones; D016297:Mutagenesis, Site-Directed; D014176:Protein Biosynthesis; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000378:metabolism; D016160:Retinoblastoma Protein / Q000378:metabolism*; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D017384:Sequence Deletion; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000096:biosynthesis
C513270:CDC37 protein, S cerevisiae; D002352:Carrier Proteins; C103269:Cdc37 protein, rat; D018797:Cell Cycle Proteins; D029721:Drosophila Proteins; D018832:Molecular Chaperones; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D016160:Retinoblastoma Protein; D029701:Saccharomyces cerevisiae Proteins; D005982:Glutathione Transferase; D001616:beta-Galactosidase