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PMID: 14576297 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Identification of peptide inhibitors of pre-mRNA splicing derived from the essential interaction domains of CDC5L and PLRG1.

Nucleic acids research ·Vol. 31 ·No. 21 ·2003-11-01 ·Pages 6104-16

Ajuh P, Lamond AI

Abstract

CDC5L and PLRG1 are both spliceosomal proteins that are highly conserved across species. They have both been shown to be part of sub- spliceosomal protein complexes that are essential for pre-mRNA splicing in yeast and humans. CDC5L and PLRG1 interact directly in vitro. This interaction is mediated by WD40 regions in PLRG1 and the C-terminal domain of CDC5L. In order to determine whether this interaction is important for the splicing mechanism, we have designed peptides corresponding to highly conserved sequences in the interaction domains of both proteins. These peptides were used in in vitro splicing experiments as competitors to the cognate sequences in the endogenous proteins. Certain peptides derived from the binding domains of both proteins were found to inhibit in vitro splicing. This splicing inhibition could be prevented by preincubating the peptides with the corresponding partner protein that had been expressed in Escherichia coli. The results from this study indicate that the interaction between CDC5L and PLRG1 is essential for pre-mRNA splicing and further demonstrate that small peptides can be used as effective splicing inhibitors.

MeSH Terms
Amino Acid Sequence Binding Sites Cell Cycle Proteins/chemistry,metabolism Cell Extracts Cell Nucleus/metabolism Conserved Sequence HeLa Cells Humans Intracellular Signaling Peptides and Proteins Molecular Sequence Data Nuclear Proteins/chemistry,metabolism Peptide Fragments/chemistry,metabolism,pharmacology Protein Binding/drug effects Protein Structure, Tertiary RNA Precursors/genetics,metabolism RNA Splicing/drug effects Sequence Alignment Spliceosomes/chemistry,drug effects,metabolism Substrate Specificity
Chemicals
Cell Cycle Proteins Cell Extracts Intracellular Signaling Peptides and Proteins Nuclear Proteins PLRG1 protein, human Peptide Fragments RNA Precursors
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Ajuh Paul
The University of Dundee, School of Life Sciences, Wellcome Trust Biocentre, Dow Street, Dundee DD1 5EH, UK.
Lamond Angus I
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gkg817
Published
2003-11-01
Pages
6104-16
Language
English
Region
England
NLM ID
0411011
PMCID
PMC275459
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/gkg817 PMC Full Text View journal details
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