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Functional domains of the yeast splicing factor Prp22p.
- Source :
-
The Journal of biological chemistry [J Biol Chem] 2001 Jun 15; Vol. 276 (24), pp. 21184-91. Date of Electronic Publication: 2001 Mar 29. - Publication Year :
- 2001
-
Abstract
- The essential Saccharomyces cerevisiae PRP22 gene encodes a 1145-amino acid DEXH box RNA helicase. Prp22p plays two roles during pre-mRNA splicing as follows: it is required for the second transesterification step and for the release of mature mRNA from the spliceosome. Whereas the step 2 function of Prp22p does not require ATP hydrolysis, spliceosome disassembly is dependent on the ATPase and helicase activities. Here we delineate a minimal functional domain, Prp22(262-1145), that suffices for the activity of Prp22p in vivo when expressed under the natural PRP22 promoter and for pre-mRNA splicing activity in vitro. The biologically active domain lacks an S1 motif (residues 177-256) that had been proposed to play a role in RNA binding by Prp22p. The deletion mutant Prp22(351-1145) can function in vivo when provided at a high gene dosage. We suggest that the segment from residues 262 to 350 enhances Prp22p function in vivo, presumably by targeting Prp22p to the spliceosome. We characterize an even smaller catalytic domain, Prp22(466-1145) that suffices for ATP hydrolysis, RNA binding, and RNA unwinding in vitro and for nuclear localization in vivo but cannot by itself support cell growth. However, the ATPase/helicase domain can function in vivo if the N-terminal region Prp22(1-480) is co-expressed in trans.
- Subjects :
- Adenosine Triphosphatases metabolism
Amino Acid Sequence
Base Sequence
Conserved Sequence
DEAD-box RNA Helicases
Fungal Proteins chemistry
Fungal Proteins genetics
Kinetics
Molecular Sequence Data
Mutagenesis, Site-Directed
Polyribonucleotides chemistry
Polyribonucleotides metabolism
RNA Splicing
RNA Splicing Factors
RNA, Messenger genetics
Recombinant Proteins chemistry
Recombinant Proteins metabolism
Sequence Alignment
Sequence Homology, Amino Acid
Spliceosomes metabolism
Substrate Specificity
Fungal Proteins metabolism
RNA Helicases metabolism
Saccharomyces cerevisiae enzymology
Saccharomyces cerevisiae genetics
Saccharomyces cerevisiae Proteins
Subjects
Details
- Language :
- English
- ISSN :
- 0021-9258
- Volume :
- 276
- Issue :
- 24
- Database :
- MEDLINE
- Journal :
- The Journal of biological chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 11283007
- Full Text :
- https://doi.org/10.1074/jbc.M101964200