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PMID: 2216708 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Uridine branch acceptor is a cis-acting element involved in regulation of the alternative processing of calcitonin/CGRP-l pre-mRNA.

Nucleic acids research ·Vol. 18 ·No. 18 ·1990-09-25 ·Pages 5365-73

Adema GJ, van Hulst KL, Baas PD

Abstract

The human calcitonin/CGRP-I (CALC-I) gene contains 6 exons and encodes two polypeptide precursors. In thyroid C-cells, calcitonin (CT) mRNA is produced by splicing of exons 1-2-3 to exon 4 (CT-encoding) and polyadenylation at exon 4. CGRP-I mRNA is produced in particular neural cells by splicing of exons 1-2-3 to exon 5 (CGRP-I-encoding) and the polyadenylated exon 6. We previously reported that model precursor RNAs containing the exon 3 to exon 5 region of the CALC-I gene are processed predominantly into CGRP-I mRNA in vitro, in nuclear extracts of several cell types (neural and non-neural). Using truncated precursor RNAs containing only the exon 3 to exon 4 region of the CALC-I gene it was shown that CT splicing is an inefficient reaction in which a uridine residue serves as the major site of lariat formation. Here we report that the low CT splicing efficiency and the dominance of CGRP-I splicing over CT splicing in vitro are primarily due to the usage of the CT-specific uridine branch acceptor. Mutation of this uridine residue into an adenosine residue resulted in a strong increase in CT splicing efficiency causing a reversal of the splicing pattern. In addition, it was shown that this point mutation also increased CT splicing efficiency in vivo. These results and data obtained from other experiments involving mutation of the CT splice acceptor site suggest that the uridine branch acceptor is a cis-acting element involved in regulation of the alternative processing of the CALC-I pre-mRNA.

MeSH Terms
Adenosine Base Sequence Blotting, Northern Calcitonin/genetics Calcitonin Gene-Related Peptide/genetics Exons Genes Humans Molecular Sequence Data Mutation RNA Precursors/genetics,metabolism RNA Splicing Uridine/metabolism
Chemicals
RNA Precursors Calcitonin Calcitonin Gene-Related Peptide Adenosine Uridine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Adema G J
Institute of Molecular Biology and Medical Biotechnology, University of Utrecht, The Netherlands.
van Hulst K L
Baas P D
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48 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/18.18.5365
Published
1990-09-25
Pages
5365-73
Language
English
Region
England
NLM ID
0411011
PMCID
PMC332211
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/18.18.5365 PMC Full Text View journal details
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