The +1,506 (A>C) mutation in the 3' untranslated region affects β-globin expression.
The 3' untranslated region (3'UTR) is known to be important to mRNA stability but the stabilization mechanism on the β-globin gene is not fully elucidated. We speculated in our previous report that +1,506 (A>C) mutation (HGVS nomenclature: *32A>C) on the β-globin 3'UTR causes β-thalassemia (β-thal) in order to destabilize the mRNA. To investigate further, we studied the expression efficiency for the mutation with a luciferase assay. We made recombinant pGL4.74 vectors in which the luciferase 3'UTR was replaced with the wild-type and mutant 3'UTR of the β-globin gene. For a comparison experiment, recombinant vectors were made not only for this mutation but also six other mutations in the β-globin 3'UTR which bring about β-thal or affect mRNA stability. The +1,506 mutation led to a 30.0% lower protein expression than normal in this assay. We concluded that this mutation destabilizes mRNA and consequently decreases the β-globin amount to finally cause β-thal. Our study highlights the crucial area of β-globin 3'UTR for protein expression.
Faculty of Health Sciences, Yamaguchi University Graduate School of Medicine, Ube, Japan. hinominako@gmail.com
Minako
M
Hino
Hitomi
H
Ito
Yasuhiro
Y
Yamashiro
Yukio
Y
Hattori
Takenori
T
Nitta
Chris
C
Adhiyanto
D020413:3' Untranslated Regions / Q000235:genetics*; D001483:Base Sequence; D045744:Cell Line, Tumor; D005786:Gene Expression Regulation; D006801:Humans; D020014:K562 Cells; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D009690:Nucleic Acid Conformation; D020871:RNA Stability; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D055544:beta-Globins / Q000737:chemistry / Q000235:genetics*; D017086:beta-Thalassemia / Q000235:genetics
D020413:3' Untranslated Regions; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D055544:beta-Globins; D008156:Luciferases