Elucidation of the expression control mechanism of blood group D antigen

2020 Fiscal Year Final Research Report

• Project/Area Number: 19K17870
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 54010:Hematology and medical oncology-related
• Research Institution: Kyorin University
• Principal Investigator: MISHIMA YUKO 杏林大学, 保健学部, 助教 (90815771)
• Project Period (FY): 2019-04-01 – 2021-03-31
• Keywords: RhD / 発現制御メカニズム

Outline of Final Research Achievements

The expression changes of RhD protein caused by the alternative splicing were examined by Western blotting using the forced expression models. Large complexes of 300 kDa or more containing RhD protein were detected more strongly than the RhD monomer. There were no differences on the expression levels among the full-length RhD protein and splicing variants that lacked exons 7-9 in various patterns. The results of immunoprecipitation of each RhD isoform and the forced expression protein of ankyrin, protein 42, spectrinα, β, and band3, which are expected to interact with RhD, showed that band3 is the only molecule that bound to RhD. However, there were no apparent differences in the binding ability among the full-length RhD protein and the other splicing variants.

Free Research Field

輸血

Academic Significance and Societal Importance of the Research Achievements

遺伝子変異の表現型からRhDとの相互作用が想定されている分子のうち、band3とRhDが直接結合することを実験的に検証しえた。一方RhDのスプライシングバリアントのmRNAレベル、蛋白レベルの発現量に差がなく、かつband3とRhDのスプライシングバリアントの結合能にも差を認めなかったことから、RhDの発現制御には選択的スプライシングのほか、未知の結合分子を含めたより複雑な分子間ネットワークが関与することが示唆された。