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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