Nucleic Acids Research, 1990, Vol. 18, No. 2 337-343
© 1990
MOLECULAR BIOLOGY |
Requirement of protein co-factor for the DNA-binding function of the human ski proto-oncogene product
1Laboratory of Molecular Genetics, Tsukuba life Science Center, Institute of Physical and Chemical Research (RIKEN) Tsukuba, lbaraki 305, Japan 2Department of Pharmaceutical Sciences, Science University of Tokyo Tokyo, Shinjuku-ku, Tokyo 162, Japan 3Molecular Oncology Laboratory, Nippon Veterinary and Zootechnical College College, Taito-ku, Tokyo 110, Japan
Received September 8, 1989. Revised December 8, 1989. Accepted December 8, 1989.
We identified the human c-ski gene product (c-Ski) as a protein with the apparent molecular weight of 100c-ski, by using a c-Ski-specific polyclonal antibody. p100C-Ski was a nuclear protein and p100C-Ski in nuclear extracts of Molt4 cells bound to calf thymus DNA cellulose, but the bacterially synthesized c-Ski did not, suggesting that Ski was associated with another protein(s) and that the Ski complex had DNA-binding activity. This hypothesis was supported by the finding that the bacterially synthesized Ski bounds to DNA cellulose after being mixed with a nuclear extract of Molt4 cells. By use of a series of deletion mutants of Ski synthesized in an in vitro translation system, two portions in Ski were found to be necessary for the DNA binding of the Ski complex: the N-proximal portion containing a cysteinlhistidine-rich domain and the Cterminal portion including a region rich in basic amino acids.
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