Nucleic Acids Research, 1994, Vol. 22, No. 15 2938-2944
© 1994
MOLECULAR BIOLOGY |
Site-directed mutagenesis study on DNA binding regions of the mouse homologue of Suppressor of Hairless, RBP-Jx
Department of Medical Chemistry, Kyoto University Faculty of Medicine Yoshida, Sakyo-ku, Kyoto 606, Japan
*To whom correspondence should be addressed
Received May 24, 1994. Accepted June 24, 1994.
To map regions important for DNA binding of the mouse homologue of Suppressor of Hairless or RBPJx protein, mutated mouse RBP-Jx cDNAs were made by insertion of oligonucleotide linkers or base replacement. DNA binding assays using the mutated proteins expressed in COS cells showed that various mutations between 218 Arg and 227 Arg decreased the DNA binding activity drastically. The DNA binding activity was not affected by ami no acid replacements within the integrase motif of the RBP-Jx protein (230His-269His). Replacements between 291 Arg and 323Tyr affected the DNA binding activity slightly but reproducibly. These results indicate that the region encompassing 218Arg-227Arg is critical for the DNA binding activity of RBP-Jx. This region did not show any significant homology to motifs or domains of the previously described DNA binding proteins. Using a truncation mutant protein RBP-Jx was shown to associate with DNA as a monomer.
+Present address: Nara Institute of Science and Technology, 8916-5, Takayama, Ikoma, Xara 630-01, Japar.
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