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Molecular and Cellular Biology, November 2003, p. 7887-7901, Vol. 23, No. 21
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.21.7887-7901.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

In Vivo Evidence that Defects in the Transcriptional Elongation Factors RPB2, TFIIS, and SPT5 Enhance Upstream Poly(A) Site Utilization{dagger}

Yajun Cui and Clyde L. Denis*

Department of Biochemistry and Molecular Biology, University of New Hampshire, Durham, New Hampshire 03824

Received 2 May 2003/ Returned for modification 26 June 2003/ Accepted 21 July 2003

While a number of proteins are involved in elongation processes, the mechanism for action of most of these factors remains unclear primarily because of the lack of suitable in vivo model systems. We identified in yeast several genes that contain internal poly(A) sites whose full-length mRNA formation is reduced by mutations in RNA polymerase II subunit RPB2, elongation factor SPT5, or TFIIS. RPB2 and SPT5 defects also promoted the utilization of upstream poly(A) sites for genes that contain multiple 3' poly(A) signaling sequences, supporting a role for elongation in differential poly(A) site choice. Our data suggest that elongation defects cause increased transcriptional pausing or arrest that results in increased utilization of internal or upstream poly(A) sites. Transcriptional pausing or arrest can therefore be visualized in vivo if a gene contains internal poly(A) sites, allowing biochemical and genetic study of the elongation process.

* Corresponding author. Mailing address: Department of Biochemistry and Molecular Biology, Rudman Hall, University of New Hampshire, Durham, NH 03824. Phone: (603) 862-2427. Fax: (603) 862-4013. E-mail: cldenis{at}cisunix.unh.edu.

{dagger} Scientific contribution no. 2087 from the New Hampshire Agriculture Experiment Station.

Molecular and Cellular Biology, November 2003, p. 7887-7901, Vol. 23, No. 21
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.21.7887-7901.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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