title: Brr2p carboxy-terminal Sec63 domain modulates Prp16 splicing RNA helicase.
abstract: RNA helicases are essential for virtually all cellular processes, however, their regulation is poorly understood. The activities of eight RNA helicases are required for pre-mRNA splicing. Amongst these, Brr2p is unusual in having two helicase modules, of which only the amino-terminal helicase domain appears to be catalytically active. Using genetic and biochemical approaches, we investigated interaction of the carboxy-terminal helicase module, in particular the carboxy-terminal Sec63-2 domain, with the splicing RNA helicase Prp16p. Combining mutations in BRR2 and PRP16 suppresses or enhances physical interaction and growth defects in an allele-specific manner, signifying functional interactions. Notably, we show that Brr2p Sec63-2 domain can modulate the ATPase activity of Prp16p in vitro by interfering with its ability to bind RNA. We therefore propose that the carboxy-terminal helicase module of Brr2p acquired a regulatory function that allows Brr2p to modulate the ATPase activity of Prp16p in the spliceosome by controlling access to its RNA substrate/cofactor.
authors.affiliation: Wellcome Trust Centre for Cell Biology, University of Edinburgh, King's Buildings, Mayfield Road, Edinburgh, EH9 3BF, UK IBPC, CNRS FRE 3630, 13, rue Pierre & Marie Curie, 75005 Paris, France jbeggs@ed.ac.uk.
authors.forename: Olivier
authors.initials: O
authors.lastname: Cordin
authors.affiliation: Wellcome Trust Centre for Cell Biology, University of Edinburgh, King's Buildings, Mayfield Road, Edinburgh, EH9 3BF, UK.
authors.forename: Daniela
authors.initials: D
authors.lastname: Hahn
authors.affiliation: Wellcome Trust Centre for Cell Biology, University of Edinburgh, King's Buildings, Mayfield Road, Edinburgh, EH9 3BF, UK.
authors.forename: Ross
authors.initials: R
authors.lastname: Alexander
authors.affiliation: Wellcome Trust Centre for Cell Biology, University of Edinburgh, King's Buildings, Mayfield Road, Edinburgh, EH9 3BF, UK.
authors.forename: Amit
authors.initials: A
authors.lastname: Gautam
authors.affiliation: Institut Pasteur, CNRS UMR3525, 25-28 rue du docteur Roux, 75015 Paris, France.
authors.forename: Cosmin
authors.initials: C
authors.lastname: Saveanu
authors.affiliation: Wellcome Trust Centre for Cell Biology, University of Edinburgh, King's Buildings, Mayfield Road, Edinburgh, EH9 3BF, UK.
authors.forename: J David
authors.initials: JD
authors.lastname: Barrass
authors.affiliation: Wellcome Trust Centre for Cell Biology, University of Edinburgh, King's Buildings, Mayfield Road, Edinburgh, EH9 3BF, UK jbeggs@ed.ac.uk.
authors.forename: Jean D
authors.initials: JD
authors.lastname: Beggs
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D000483:Alleles; D053487:DEAD-box RNA Helicases / Q000378:metabolism; D006358:Hot Temperature; D007438:Introns; D009154:Mutation; D017434:Protein Structure, Tertiary; D012313:RNA / Q000378:metabolism; D020365:RNA Helicases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012326:RNA Splicing; D000072260:RNA Splicing Factors; D017411:Ribonucleoproteins, Small Nuclear / Q000737:chemistry; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D000072260:RNA Splicing Factors; D017411:Ribonucleoproteins, Small Nuclear; C545711:SNRNP200 protein, human; D029701:Saccharomyces cerevisiae Proteins; D012313:RNA; D000251:Adenosine Triphosphatases; C104158:BRR2 protein, S cerevisiae; C075669:PRP16 protein, S cerevisiae; C066490:PRP2 protein, S cerevisiae; D053487:DEAD-box RNA Helicases; D020365:RNA Helicases