Characterization of the Schizosaccharomyces pombe Spt5-Spt4 complex

• DOI: 10.1261/rna.1572709
• Publication Date: 2009-05-22T00:54:35Z
• Authors: Beate Schwer; Susanne Schneider; Yi Pei; Anna Aronova; Stewart Shuman
• Keywords: Schizosaccharomyces

The Spt5-Spt4 complex regulates early transcription elongation by RNA polymerase II and has an imputed role in pre-mRNA processing via its physical association with mRNA capping enzymes. Here we characterize the Schizosaccharomyces pombe core as a heterodimer map trypsin-resistant Spt4-binding domain within Spt5 subunit. A genetic analysis of Spt4 S. revealed it to be inessential for growth at 25°C–30°C but critical 37°C. These results echo conditional spt4 Δ phenotype budding yeast, where find that Saccharomyces cerevisiae are functionally interchangeable. Complementation two-hybrid assay Spt4-Spt5 interaction provided readout effects 33 missense truncation mutations on function vivo, which were interpreted light recent crystal structure fused fragment Spt5. Our highlight importance Zn 2+ -binding residues—Cys12, Cys15, Cys29, Asp32—and Ser57, conserved constituent interface. 990-amino acid protein exceptionally regular carboxyl-terminal (CTD) composed 18 nonapeptide repeats. We few three nonamer repeats sufficed growth, only when was present. Synthetic lethality spt5 1-835 double mutant 34°C suggests central overlaps CTD.