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Molecular and Cellular Biology, March 2003, p. 1961-1967, Vol. 23, No. 6
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.6.1961-1967.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

RNA Polymerase II Accumulation in the Promoter-Proximal Region of the Dihydrofolate Reductase and {gamma}-Actin Genes

Chonghui Cheng1 and Phillip A. Sharp1,2,3*

Center for Cancer Research,1 Department of Biology,2 McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, Massachusetts3

Received 23 July 2002/ Returned for modification 4 September 2002/ Accepted 19 December 2002

The carboxyl-terminal domain (CTD) of RNA polymerase II (Pol II) can be phosphorylated at serine 2 (Ser-2) and serine 5 (Ser-5) of the CTD heptad repeat YSPTSPS, and this phosphorylation is important in coupling transcription to RNA processing, including 5' capping, splicing, and polyadenylation. The mammalian endogenous dihydrofolate reductase and {gamma}-actin genes have been used to study the association of Pol II with different regions of transcribed genes (promoter-proximal compared to distal regions) and the phosphorylation status of its CTD. For both genes, Pol II is more concentrated in the promoter-proximal regions than in the interior regions. Moreover, different phosphorylation forms of Pol II are associated with distinct regions. Ser-5 phosphorylation of Pol II is concentrated near the promoter, while Ser-2 phosphorylation is observed throughout the gene. These results suggest that the accumulation of paused Pol II in promoter-proximal regions may be a common feature of gene regulation in mammalian cells.

* Corresponding author. Mailing address: Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139-4307. Phone: (617) 253-6421. Fax: (617) 253-3867. E-mail: sharppa{at}mit.edu.

Molecular and Cellular Biology, March 2003, p. 1961-1967, Vol. 23, No. 6
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.6.1961-1967.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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