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Protein Interaction of Retinoblastoma Gene Product pRb110 with M_r 73,000 Heat Shock Cognate Protein¹

Departments of Pathology [T.N., S.T., N.S., K.K.] and of Obstetrics and Gynecology [T.N., S.S.], Sapporo Medical College, S. I. W. 17, Chuo-ku, 060 Sapporo, Japan

Both the tumor suppressor gene products, the retinoblastoma sensitivity gene product pRb110 and p53, are found in oligomer complexes with the oncogene products of the DNA tumor viruses. It has been demonstrated that p53 binds to the M_r 70,000 heat shock protein family. However, the protein association of pRb110 with the M_r 70,000 heat shock protein family is not yet known. We analyzed the immunoprecipitates made with TYK-nu human ovarial carcinoma cell lysate and anti-pRb110 or anti-heat shock protein monoclonal antibodies. In this paper, we demonstrate that pRb110 is associated with the M_r 73,000 heat shock cognate protein, but not with the M_r 72,000 heat shock protein. This selective protein association was also detected in HeLa cervical carcinoma cells. Furthermore, the protein complexes of the M_r 73,000 heat shock cognate protein and pRb110 were dissociated with the presence of ATP, but not with ADP and the nonhydrolyzable ATP analogue, ATPS. This indicates that the dissociation is dependent on the ATP hydrolysis. These data may suggest an as yet undefined important role of M_r 73,000 heat shock cognate protein in the cell growth control in collaboration with pRb110.

¹ This work was supported by a Grant-in-Aid from the Ministry of Education, Culture and Science of Japan.

² To whom requests for reprints should be addressed.

Received 9/25/92. Accepted 1/15/93.

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