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Molecular and Cellular Biology, November 2001, p. 7617-7628, Vol. 21, No. 22
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.22.7617-7628.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

The Transcription Elongation Factor CA150 Interacts with RNA Polymerase II and the Pre-mRNA Splicing Factor SF1

Aaron C. Goldstrohm,1 Todd R. Albrecht,1 Carles Suñé,1,dagger Mark T. Bedford,2 and Mariano A. Garcia-Blanco1,3,4,*

Departments of Genetics,1 Microbiology,3 and Medicine,4 Duke University Medical Center, Durham, North Carolina 27710, and Department of Carcinogenesis, MD Anderson Cancer Center, University of Texas, Smithville, Texas 789572

Received 17 May 2001/Returned for modification 20 July 2001/Accepted 17 August 2001

CA150 represses RNA polymerase II (RNAPII) transcription by inhibiting the elongation of transcripts. The FF repeat domains of CA150 bind directly to the phosphorylated carboxyl-terminal domain of the largest subunit of RNAPII. We determined that this interaction is required for efficient CA150-mediated repression of transcription from the alpha 4-integrin promoter. Additional functional determinants, namely, the WW1 and WW2 domains of CA150, were also required for efficient repression. A protein that interacted directly with CA150 WW1 and WW2 was identified as the splicing-transcription factor SF1. Previous studies have demonstrated a role for SF1 in transcription repression, and we found that binding of the CA150 WW1 and WW2 domains to SF1 correlated exactly with the functional contribution of these domains for repression. The binding specificity of the CA150 WW domains was found to be unique in comparison to known classes of WW domains. Furthermore, the CA150 binding site, within the carboxyl-terminal half of SF1, contains a novel type of proline-rich motif that may be recognized by the CA150 WW1 and WW2 domains. These results support a model for the recruitment of CA150 to repress transcription elongation. In this model, CA150 binds to the phosphorylated CTD of elongating RNAPII and SF1 targets the nascent transcript.

* Corresponding author. Mailing address: Department of Genetics, Duke University Medical Center, Box 3053, Durham, NC 27710. Phone: (919) 613-8632. Fax: (919) 613-8646. E-mail: garci001{at}mc.duke.edu.

dagger Present address: Institute of Medical Microbiology, 4003 Basel, Switzerland.

Molecular and Cellular Biology, November 2001, p. 7617-7628, Vol. 21, No. 22
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.22.7617-7628.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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