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Molecular and Cellular Biology, September 1999, p. 6154-6163, Vol. 19, No. 9
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

MCM Proteins Are Associated with RNA Polymerase II Holoenzyme

K. Yankulov,1,* I. Todorov,2 P. Romanowski,3 D. Licatalosi,4 K. Cilli,4 S. McCracken,5 R. Laskey,3 and D. L. Bentley4

Department of Molecular Biology and Genetics, University of Guelph, Guelph, Ontario N1G 2W1,1 and Banting & Best Department of Medical Research, University of Toronto, Toronto, Ontario M5G 1L6,5 Canada; Department of Biochemistry and Molecular Genetics, University of Colorado Health Sciences Center, Denver, Colorado 802624; Desmos Inc., San Diego, California 921212; and Wellcome/CRC Institute, Cambridge CB2 1OR, United Kingdom3

Received 20 April 1999/Returned for modification 26 May 1999/Accepted 1 June 1999

MCMs are a family of proteins related to ATP-dependent helicases that bind to origin recognition complexes and are required for initiation of DNA replication. We report that antibodies against MCM2(BM28) specifically inhibited transcription by RNA polymerase II (Pol II) in microinjected Xenopus oocytes. Consistent with this observation, MCM2 and other MCMs copurified with Pol II and general transcription factors (GTFs) in high-molecular-weight holoenzyme complexes isolated from Xenopus oocytes and HeLa cells. Pol II and GTFs also copurified with MCMs isolated by anti-MCM3 immunoaffinity chromatography. MCMs were specifically displaced from the holoenzyme complex by antibody against the C-terminal domain (CTD) of Pol II. In addition, MCMs bound to a CTD affinity column, suggesting that their association with holoenzyme depends in part on this domain of Pol II. These results suggest a new function for MCM proteins as components of the Pol II transcriptional apparatus.

* Corresponding author. Mailing address: Department of Molecular Biology and Genetics, University of Guelph, Guelph, Ontario N1G 2W1, Canada. Phone: (519) 824-4120, ext. 6466. Fax: (519) 837-2075. E-mail: yankulov{at}uoguelph.ca.

Molecular and Cellular Biology, September 1999, p. 6154-6163, Vol. 19, No. 9
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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