XXIth YGM Conference
Göteborg, Sweden
July 7-12th, 2003

Conference Web Site ( http://www.yeast2003.se )

Abstract 7-31 Iwr1: A novel, conserved RNAPII-interacting protein implicated in transcriptional elongation in Saccharomyces cerevisiae and Drosophila melanogaster.
Nira Datta (1), Nevan Krogan (1), Gerard Cagney (1), Andrew Emili (1), Jack Greenblatt (1), Ali Shilatifard (2), Steve Buratowski (3)
(1) Medical Genetics, University of Toronto, 112 College Street, Toronto, M5G 1L6, Canada (niradatta@hotmail.com); (2) St. Louis University, Biochemistry Room 157, 1402 S. Grand Blvd., St. Louis, MO, 63104, USA; (3) Harvard Medical School, Department of Biological Chemistry and Molecular Pharmacology, Bldg. C1, Room 210, 240 Longwood Avenue, Boston, MA 02115, USA

In order to discover novel factors involved in elongation by RNA polymerase II (RNAPII), several subunits of S. cerevisiae RNAPII were TAP-tagged and purified. One novel factor identified by mass spectrometry that co-purified with RNAPII is the previously uncharacterized ORF, Ydl115c, which we have named Iwr1 (interacts with RNA polymerase II). When Iwr1 was tagged and purified, it co-purified with a nearly stoichiometric amount of transcriptionally active RNAPII. An iwr1 deletion strain is sensitive to 6-azauracil, suggesting Iwr1 could have a role in transcriptional elongation. In synthetic genetic array (SGA) analysis, an iwr1 deletion was found to genetically interact with several known elongation factors, including TFIIS and components of the Paf1 complex, with mediator subunits Gal11 and Soh1, and with components of the Set1 and Set3 complexes. Interestingly, iwr1 also results in synthetic lethality when combined with a deletion of RPB9, a subunit of RNAPII previously implicated in transcriptional elongation. Finally, antibody was generated against the D. melanogaster homologue of Iwr1. dIwr1 localizes to actively transcribed regions on polytene chromosomes: it co-localizes with RNAPII phosphorylated on Ser2 of the CTD, a form of polymerase known to be involved in transcriptional elongation. Our combined data suggest that Iwr1 is an evolutionarily conserved protein which functions in transcriptional elongation.


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