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PMID: 10393914 Published · ppublish English Journal Article

Single-nucleotide polymorphisms can cause different structural folds of mRNA.

Shen LX, Basilion JP, Stanton VP

Abstract

Single-nucleotide polymorphisms (SNPs) are the most common type of genetic variation in man. Genes containing one or more SNPs can give rise to two or more allelic forms of mRNAs. These mRNA variants may possess different biological functions as a result of differences in primary or higher order structures that interact with other cellular components. Here we report the observation of marked differences in mRNA secondary structure associated with SNPs in the coding regions of two human mRNAs: alanyl tRNA synthetase and replication protein A, 70-kDa subunit (RPA70). Enzymatic probing of SNP-containing allelic fragments of the mRNAs revealed pronounced allelic differences in cleavage pattern at sites 14 or 18 nt away from the SNP, suggesting that a single-nucleotide variation can give rise to different mRNA folds. By using phosphorothioate oligodeoxyribonucleotides complementary to the region of different allelic structures in the RPA70 mRNA, but not extending to the SNP itself, we find that the SNP exerts an allele-specific effect on the accessibility of its flanking site in the endogenous human RPA70 mRNA. This further supports the allele-specific structural features identified by enzymatic probing. These results demonstrate the contribution of common genetic variation to structural diversity of mRNA and suggest a broader role than previously thought for the effects of SNPs on mRNA structure and, ultimately, biological function.

MeSH Terms
Alanine-tRNA Ligase/genetics Alleles Base Sequence DNA Helicases/genetics DNA-Binding Proteins/genetics Escherichia coli/enzymology Genetic Variation Humans Macromolecular Substances Molecular Sequence Data Mutagenesis, Site-Directed Nucleic Acid Conformation Polymerase Chain Reaction Polymorphism, Genetic RNA, Messenger/chemistry,genetics Replication Protein A Ribonuclease H/metabolism
Chemicals
DNA-Binding Proteins Macromolecular Substances RNA, Messenger RPA1 protein, human Replication Protein A Ribonuclease H DNA Helicases Alanine-tRNA Ligase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Shen L X
Variagenics, Inc., 60 Hampshire Street, Cambridge, MA 02139, USA. lshen@variagenics.com
Basilion J P
Stanton V P
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
1999-07-06
Pages
7871-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC22154
Subset
IM
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