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Nucleic Acids Research, 1990, Vol. 18, No. 16 4843-4849
© 1990

ENZYMOLOGY

A heteromeric transcription factor required for mammalian RNA polymerase II

Shigetaka Kitajima, Yujiro Tanaka, Tatsuo Kawaguchi, Takayuki Nagaoka1, Sherman M. Weissman2 and Yukio Yasukochi

Department of Human Genetics, Medical Research Institute, Tokyo Medical and Dental University 1-5-45 Yushima, Bunkyo-ku, Tokyo 113 1Department of Clinical Genetics, School of Health Sciences, Kyorin University 476 Miyashitacho, Hachioji, Tokyo 192, Japan 2Department of Human Genetics, Yale University School of Medicine 333 Cedar Street, New Haven, CT 06510-8005, USA

Received April 13, 1990. Revised July 18, 1990. Accepted July 18, 1990.

A general transcription factor, FC, essential for specific initiation of in vitro transcription by mammalian RNA polymerase II was identified and a procedure developed to purify it to near homogeneity from HeLa cell nuclei. Purified FC is composed of two polypeptides of apparent molecular masses 80 kDa and 30 kDa, on SOS-PAGE, and has a native size of 280 kDa estimated by gel filtration column. Both polypeptides were shown to be essential for reconstituting in vitro transcription activity. Biochemical analysis showed that the 80 kDa and 30 kDa components were present in a 1: 1 molar ratio. FC was also demonstrated to interact directly or indirectly with purified RNA polymerase II. Similarities between FC and transcription factors reported by others from human, rat or Drosophila cells are discussed.


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