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PMID: 9233818 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

An evolutionarily conserved U5 snRNP-specific protein is a GTP-binding factor closely related to the ribosomal translocase EF-2.

The EMBO journal ·Vol. 16 ·No. 13 ·1997-07-01 ·Pages 4092-106

Fabrizio P, Laggerbauer B, Lauber J, Lane WS, Lührmann R

Abstract

The driving forces behind the many RNA conformational changes occurring in the spliceosome are not well understood. Here we characterize an evolutionarily conserved human U5 small nuclear ribonucleoprotein (snRNP) protein (U5-116kD) that is strikingly homologous to the ribosomal elongation factor EF-2 (ribosomal translocase). A 114 kDa protein (Snu114p) homologous to U5-116kD was identified in Saccharomyces cerevisiae and was shown to be essential for yeast cell viability. Genetic depletion of Snu114p results in accumulation of unspliced pre-mRNA, indicating that Snu114p is essential for splicing in vivo. Antibodies specific for U5-116kD inhibit pre-mRNA splicing in a HeLa nuclear extract in vitro. In HeLa cells, U5-116kD is located in the nucleus and colocalizes with snRNP-containing subnuclear structures referred to as speckles. The G domain of U5-116kD/Snu114p contains the consensus sequence elements G1-G5 important for binding and hydrolyzing GTP. Consistent with this, U5-116kD can be cross-linked specifically to GTP by UV irradiation of U5 snRNPs. Moreover, a single amino acid substitution in the G1 sequence motif of Snu114p, expected to abolish GTP-binding activity, is lethal, suggesting that GTP binding and probably GTP hydrolysis is important for the function of U5-116kD/Snu114p. This is to date the first evidence that a G domain-containing protein plays an essential role in the pre-mRNA splicing process.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Caenorhabditis elegans Cell Survival Conserved Sequence Cross-Linking Reagents DNA, Complementary Evolution, Molecular GTP-Binding Proteins/chemistry,genetics,metabolism Genes, Lethal Guanosine Triphosphate/metabolism HeLa Cells Humans Mice Molecular Sequence Data Mutagenesis Peptide Elongation Factor 2 Peptide Elongation Factor G Peptide Elongation Factors/chemistry Phenotype RNA Precursors RNA Splicing Ribonucleoprotein, U5 Small Nuclear/chemistry,genetics,metabolism Subcellular Fractions Ultraviolet Rays
Chemicals
Cross-Linking Reagents DNA, Complementary Peptide Elongation Factor 2 Peptide Elongation Factor G Peptide Elongation Factors RNA Precursors Ribonucleoprotein, U5 Small Nuclear Guanosine Triphosphate GTP-Binding Proteins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Fabrizio P
Institut für Molekularbiologie und Tumorforschung, Philipps-Universität Marburg, Germany.
Laggerbauer B
Lauber J
Lane W S
Lührmann R
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/16.13.4092
Published
1997-07-01
Pages
4092-106
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1170032
Subset
IM
Databases
GENBANK
U97079
External Links
PubMed source DOI: 10.1093/emboj/16.13.4092 PMC Full Text View journal details
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