Identification and characterization of Prp45p and Prp46p, essential pre-mRNA splicing factors.
Through exhaustive two-hybrid screens using a budding yeast genomic library, and starting with the splicing factor and DEAH-box RNA helicase Prp22p as bait, we identified yeast Prp45p and Prp46p. We show that as well as interacting in two-hybrid screens, Prp45p and Prp46p interact with each other in vitro. We demonstrate that Prp45p and Prp46p are spliceosome associated throughout the splicing process and both are essential for pre-mRNA splicing. Under nonsplicing conditions they also associate in coprecipitation assays with low levels of the U2, U5, and U6 snRNAs that may indicate their presence in endogenous activated spliceosomes or in a postsplicing snRNP complex.
Albers
Michael
M
Wellcome Trust Centre for Cell Biology, University of Edinburgh, UK.
Diment
Ann
A
Muraru
Mariela
M
Russell
Caroline S
CS
Beggs
Jean D
JD
D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D017931:DNA Primers; D011233:Precipitin Tests; D011485:Protein Binding; D012326:RNA Splicing; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
D017931:DNA Primers; D029701:Saccharomyces cerevisiae Proteins