2000 Fiscal Year Final Research Report Summary
Gene expression mechanism in normal and red cell membrane disorders
| Project/Area Number |
11670151
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Pathological medical chemistry
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| Research Institution | KAWASAKI MEDICAL SCHOOL |
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Principal Investigator |
YAMADA Osamu Kawasaki Medical School, Medicine, Associate Professor, 医学部, 助教授 (50104790)
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| Co-Investigator(Kenkyū-buntansha) |
KAKU Mayumi Kawasaki Medical School, Medicine, Research Associate, 医学部, 助手 (20319940)
WADA Hideho Kawasaki Medical School, Medicine, Assistant Professor, 医学部, 講師 (70191830)
YAWATA Yoshihito Kawasaki Medical School, Medicine, Professor, 医学部, 教授 (70069011)
MIKAMI Makoto Kawasaki Medical School, Medicine, Research Associate, 医学部, 助手 (90309550)
SUETSUGU Keisyu Kawasaki Medical School, Medicine, Research Associate, 医学部, 助手 (60309549)
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| Project Period (FY) |
1999 – 2000
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| Keywords | red cell membrane abnormality / Hereditary spherocytosis / gene expression / b-spectrin / band3 / protein 4.2 / 5'-CpG-3' sites / methylation |
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| Research Abstract |
It is known that major membrane proteins of the erythroid cells are orderly expressed in a cascade fashion. Therefore, the control mechanism of the gene expresson of these proteins should exist. A possible abnormality of this control mechanism is assumed to be one of the the factors pathognomonic for hereditary spherocytosis (HS). From our results by previous investigations, it became evident that some of HS patients do not manifest the deficiency of membrane proteins, and that the abnormal expression of membrane protein genes may be crucial for disease states. The states of methylation of the 5'-CpG-3' sites are now known to regulate promoter functions by modifying the extent of gene expression. Therefore, the metylation profiles were examined by the bisulfite genomic sequencing method in the genomic DNAs of the human erythroid membrane proteins ; protein 4.2 gene, band 3 gene, and β-spectrin gene.
1) The 5'-CpG-3' sites at the promoter regions of protein 4.2 and band 3 genes obtained from normal human peripheral blood mononuclear cells were extensively methylated, contrary to the fact that the promoter of β-spectrin gene was totally umethylated. 2) During erythroid differentiation, protein 4.2 gene was unmethylated in DNAs from the cell line UT-7/EPO, but became methylated in cultured erythroblasts from peripheral BFU-E.3) We also performed gene analyses for the ankyrin gene mutations in HS patients. As the result, we found 4 frameshifts, 2 nonsense mutations and 3 abnormal splicings as the etiologic abnormal gene mutations. Characteristically, missense mutations were rare, and the hot spots of these mutations were not present in the ankyrn gene.
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