TY - JOUR
T1 - Dissociation of DNA binding and in vitro transcriptional activities dependent on the C terminus of p53 proteins
AU - Kaku, Shinsuke
AU - Albor, Amador
AU - Kulesz-Martin, Molly
N1 - Funding Information:
We thank Dr. Danny Reinberg and Dr. Ron Drapkin for supplying the plasmids used to construct the p53-dependent transcription templates and for helpful discussions and Dr. Stephen Friend for providing the human p53 expression plasmid. We are grateful to Dr. Enrico Mihich and to Dr. Takehiro Yamagishi and Dr. Shiro Nakaike of Taisho Pharmaceutical Co., Ltd. for scientific interactions and support. Thanks also to Jennifer Carlson and Michelle Bryant for assistance with preparation of this manuscript. This work was supported by the Taisho Pharmaceutical Co., Ltd., and RPCI Institute Core Grant CA16056.
PY - 2001
Y1 - 2001
N2 - Wild type p53 protein requires posttranslational modification within a carboxy-terminal negative regulatory domain to activate DNA binding and transcription. Binding of monoclonal antibody PAb421 to the carboxy-terminal domain reproduces this activation. In the absence of PAb421, we found that wild type p53 bound actively to a template containing two copies of the p21WAF1 p53 binding site. However, in an in vitro transcription assay with partially purified basal transcription factors, p53 only partially activated transcription from the same binding site and required PAb421 for full activation. Oncogenic missense mutant p53 proteins (N239 to S239, G245 to S245, R273 to H273) bound the WAF1 doublet significantly and were activated further by PAb421. However, these mutants were inactive in the transcription assay, even with PAb421. These results indicate that sequence-specific binding and transcriptional activities of p53 can be dissociated through C-terminal interactions and suggest that conformational changes induced by the mutations alter p53 interactions with basal transcription factors.
AB - Wild type p53 protein requires posttranslational modification within a carboxy-terminal negative regulatory domain to activate DNA binding and transcription. Binding of monoclonal antibody PAb421 to the carboxy-terminal domain reproduces this activation. In the absence of PAb421, we found that wild type p53 bound actively to a template containing two copies of the p21WAF1 p53 binding site. However, in an in vitro transcription assay with partially purified basal transcription factors, p53 only partially activated transcription from the same binding site and required PAb421 for full activation. Oncogenic missense mutant p53 proteins (N239 to S239, G245 to S245, R273 to H273) bound the WAF1 doublet significantly and were activated further by PAb421. However, these mutants were inactive in the transcription assay, even with PAb421. These results indicate that sequence-specific binding and transcriptional activities of p53 can be dissociated through C-terminal interactions and suggest that conformational changes induced by the mutations alter p53 interactions with basal transcription factors.
KW - Eukaryotic transcriptional regulation
KW - Mutant p53 proteins
KW - Transcription factors
KW - Tumor suppressor p53
UR - http://www.scopus.com/inward/record.url?scp=0034805653&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034805653&partnerID=8YFLogxK
U2 - 10.1006/bbrc.2000.4060
DO - 10.1006/bbrc.2000.4060
M3 - Article
C2 - 11162500
AN - SCOPUS:0034805653
SN - 0006-291X
VL - 280
SP - 204
EP - 211
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -