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Volume 271, Number 40, Issue of October 4, 1996 pp. 24551-24556
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

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The Retinoblastoma Gene Product (Rb) Induces Binding of a Conformationally Inactive Nuclear Factor-B

(Received for publication, May 6, 1996, and in revised form, July 18, 1996)

From the Laboratory of Molecular Virology, National Cancer Institute, Bethesda, Maryland 20892-5005

Nuclear factor-B (NF-B) regulates expression of several viral and cellular genes including the human immunodeficiency virus long terminal repeat, major histocompatibility complex class I, and interleukin 2R cytokine genes. Here we report that the retinoblastoma gene product (Rb) stimulates binding of the NF-B p50 homodimer. The addition of Rb protein to an in vitro gel shift binding assay stimulated p50 binding greater than 10-fold. Interestingly, by analyzing NF-B-dependent transcription activity in vitro, we demonstrate that Rb suppresses transcriptional activity of p50. Chymotrypsin analysis suggests that Rb induces a conformational change in the NF-B-DNA complex, resulting in binding of a transcriptionally inactive complex. Finally, we demonstrate by coimmunoprecipitation analysis that the Rb-p50 complex is present in Jurkat cell extracts. Our results suggest that Rb may play an important role in regulation of NF-B transcriptional activity.

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