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Molecular and Cellular Biology, April 2002, p. 1971-1980, Vol. 22, No. 7
0270-7306/02/$04.00+0     DOI: 10.1128/MCB.22.7.1971-1980.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Ctr9, Rtf1, and Leo1 Are Components of the Paf1/RNA Polymerase II Complex

Cherie L. Mueller and Judith A. Jaehning*

Department of Biochemistry and Molecular Genetics and Molecular Biology Program, University of Colorado Health Sciences Center, Denver, Colorado

Received 9 November 2001/ Returned for modification 11 December 2001/ Accepted 21 December 2001

The Saccharomyces cerevisiae Paf1-RNA polymerase II (Pol II) complex is biochemically and functionally distinct from the Srb-mediator form of Pol II holoenzyme and is required for full expression of a subset of genes. In this work we have used tandem affinity purification tags to isolate the Paf1 complex and mass spectrometry to identify additional components. We have established that Ctr9, Rtf1, and Leo1 are factors that associate with Paf1, Cdc73, and Pol II, but not with the Srb-mediator. Deletion of either PAF1 or CTR9 leads to similar severe pleiotropic phenotypes, which are unaltered when the two mutations are combined. In contrast, we found that deletion of LEO1 or RTF1 leads to few obvious phenotypes, although mutation of RTF1 suppresses mutations in TATA-binding protein, alters transcriptional start sites, and affects elongation. Remarkably, deletion of LEO1 or RTF1 suppresses many paf1{Delta} phenotypes. In particular, an rtf1{Delta} paf1{Delta} double mutant grew faster, was less temperature sensitive, and was more resistant to caffeine and hydroxyurea than a paf1{Delta} single mutant. In addition, expression of the G1 cyclin CLN1, reduced nearly threefold in paf1{Delta}, is restored to wild-type levels in the rtf1{Delta} paf1{Delta} double mutant. We suggest that lack of Paf1 results in a defective complex and a block in transcription, which is relieved by removal of Leo1 or Rtf1.

* Corresponding author. Mailing address: Department of Biochemistry and Molecular Genetics, Box B121, University of Colorado Health Sciences Center, 4200 E. 9th Ave., Denver, CO 80262. Phone: (303) 315-3004. Fax: (303) 315-3326. E-mail: Judith.Jaehning{at}UCHSC.edu.

Molecular and Cellular Biology, April 2002, p. 1971-1980, Vol. 22, No. 7
0022-538X/02/$04.00+0     DOI: 10.1128/MCB.22.7.1971-1980.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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