Specific interaction of pRB with a rat genomic DNA fragment, REC11.
The retinoblastoma gene product (pRB) has been known to contain a sequence-nonspecific DNA binding activity. It is unknown whether pRB can recognize and bind to specific DNA sequences. We have recently identified a rat genomic DNA fragment, termed REC11, which can interact with rat Cdc37-related protein (RCdc37). In this study, we have found that pRB could interact with the REC11, DNA in a sequence-specific manner. A series of GST-RB deletion mutants was used in the gel shift assays to define the domain of pRB responsible for this interaction. GST-RB (385-611) and GST-RB (612-928) completely lost the binding activity, while GST-RB (555-682) retained an activity to associate with the REC11 DNA, indicating that the continuous spacer region of pRB might be important for this sequence-specific DNA binding activity.
Ozaki
T
T
Division of Biochemistry, Chiba Cancer Center Research Institute, Japan.
Sakiyama
S
S
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D017354:Point Mutation; D011485:Protein Binding; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D016160:Retinoblastoma Protein / Q000235:genetics / Q000378:metabolism*
D011993:Recombinant Fusion Proteins; D016160:Retinoblastoma Protein; D004247:DNA; D005982:Glutathione Transferase