Abstract
The G20210A polymorphism has been shown to alter the efficiency of prothrombin mRNA processing. Here we show that the G20210A mutation also alters prothrombin mRNA stability. Three-fold more prothrombin protein and mRNA were produced in NIH-3T3 cells transfected with the prothrombin cDNAs containing the 20210A variant compared to cells expressing the 20210G variant. mRNA stability assays using chimeric globin transcripts harboring the G or A variant of the 97 nt prothrombin 3'-UTR indicated that the 20210G variant conferred greater instability to the globin reporter transcript than the A variant in transfected HepG2 cells. Both variants of the prothrombin 3'-UTR were shown to provide binding sites for a number of cellular proteins including HuR, an RNA binding protein associated with mRNA stability. Our results indicate that the G20210A is a bifunctional polymorphism, as it not only alters the efficiency of mRNA processing, but also the decay rate of prothrombin mRNA.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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3' Untranslated Regions / genetics*
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3T3 Cells
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Alleles
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Animals
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Antigens, Surface*
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Binding Sites
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DNA, Complementary / genetics
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DNA, Recombinant / genetics
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ELAV Proteins
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ELAV-Like Protein 1
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Electrophoretic Mobility Shift Assay
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Gene Expression Regulation
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Genes, Reporter
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Globins / genetics
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Heterogeneous Nuclear Ribonucleoprotein D0
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Heterogeneous-Nuclear Ribonucleoprotein D*
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Humans
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Mice
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Polymorphism, Genetic*
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Protein Biosynthesis
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Prothrombin / biosynthesis
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Prothrombin / genetics*
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RNA, Messenger / genetics
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RNA, Messenger / metabolism*
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RNA-Binding Proteins / metabolism
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Regulatory Sequences, Nucleic Acid
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Transcription, Genetic
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Transfection
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Tumor Cells, Cultured
Substances
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3' Untranslated Regions
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Antigens, Surface
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DNA, Complementary
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DNA, Recombinant
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ELAV Proteins
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ELAV-Like Protein 1
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ELAVL1 protein, human
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HNRNPD protein, human
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Heterogeneous Nuclear Ribonucleoprotein D0
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Heterogeneous-Nuclear Ribonucleoprotein D
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Hnrpd protein, mouse
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RNA, Messenger
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RNA-Binding Proteins
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Prothrombin
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Globins