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PMID: 17875666 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Ntr1 activates the Prp43 helicase to trigger release of lariat-intron from the spliceosome.

Genes & development ·Vol. 21 ·No. 18 ·2007-09-15 ·Pages 2312-25

Tanaka N, Aronova A, Schwer B

Abstract

DEAD/H-box NTPases remodel the spliceosome at multiple steps during the pre-mRNA splicing cycle. The RNA-dependent NTPase Prp43 catalyzes dissociation of excised lariat-intron from the spliceosome, but it is unclear how Prp43 couples the energy of ATP hydrolysis to intron release. Here, we report that activation of Prp43's inherently feeble helicase activity by the splicing factor Ntr1 is required for lariat-intron release. Lethal Prp43 mutants T384A and T384V, which are active for ATP hydrolysis and fail to dissociate lariat-intron from spliceosomes, are refractory to stimulation of RNA unwinding by Ntr1. An N-terminal 120-amino-acid segment of Ntr1 suffices for binding to Prp43 and for stimulating its helicase activity. We identify missense mutations in Prp43 and Ntr1 that disrupt protein-protein interaction and impair Ntr1 enhancement of Prp43 RNA unwinding. Our results demonstrate for the first time that regulating the motor activity of a DEAH-box protein by an accessory factor is critical for mRNA splicing.

MeSH Terms
Amino Acid Sequence DEAD-box RNA Helicases/genetics,metabolism Enzyme Activation Introns Molecular Sequence Data Mutant Proteins/metabolism Mutation, Missense Protein Binding Protein Structure, Tertiary RNA Splicing Saccharomyces cerevisiae Proteins/chemistry,genetics,metabolism,physiology Sequence Homology, Amino Acid Spliceosomes/metabolism
Chemicals
Mutant Proteins Saccharomyces cerevisiae Proteins Spp382 protein, S cerevisiae PRP43 protein, S cerevisiae DEAD-box RNA Helicases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Tanaka Naoko
Department of Microbiology and Immunology, Weill Cornell Medical College, New York, NY 10021, USA.
Aronova Anna
Schwer Beate
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45 references, click to expand
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.1580507
Published
2007-09-15
Pages
2312-25
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC1973145
Subset
IM
Grants
NIGMS NIH HHS · R01 GM050288 · United States
NIGMS NIH HHS · GM50288 · United States
External Links
PubMed source DOI: 10.1101/gad.1580507 PMC Full Text View journal details
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