| Project/Area Number |
02807156
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Obstetrics and gynecology
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| Research Institution | Tokai University School of Medicine |
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Principal Investigator |
IWASAKI Katsuhiko Tokai University School of Medicine, Obstetrics and Gynecology, Associate Professor, 医学部, 助教授 (10119646)
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| Co-Investigator(Kenkyū-buntansha) |
MORIUCHI Tetauya Tokai University School of Medicine, Cell Biology, Associate Professor, 医学部, 助教授 (20174394)
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| Project Period (FY) |
1990 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1991: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1990: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | PCNA / DNA polymerase / congenital anomalies / VNTR / PCNA / DNAポリメラ-ゼβ / γークリスタリン / DNA複製遺伝子 |
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| Research Abstract |
High molecular weight DNAS were purified from the placentas or autopsied tissues of severe congenital malformations. Fifteen DNA samples were examined for the abnormal restriction fragments by Southern blot analysis using cDNAs for DNA polymerase-alpha, DNA polymerase-beta and DNA polymerase-delta auxiliary protein(PCNA)as probes. The DNA samples were digested with 10 restriction endonucleases. In one case of congenital malformations, the 3 probes detected abnormal restriction fragments in the DNA digests of 3-5 restriction enzymes, respectively. All abnormal bands were seen heterozygously. Clinical examination of the patient(MY)revealed microcephalia, urogenital anomaly, arthrogryposis and scoliosis. In the other 14 DNA samples, no abnormal bands were detected.
DNA polymerase delta auxiliary protein(PCNA)is an essential element for DNA replication in eukaryotes and the gene is highly conserved throughout the plant and animal kingdoms(Suzuka et al. PNAS 1989). Yamaguchi et al. (MCB 1990)demonstrated that Drosophila HOX proteins specifically bind to the 5'-flanking region of Drosophila PCNA gene. In the same region of the human PCNA gene, a perfect octamer motif ATTTGCAT is found. These observations strongly suggest that the expression of PCNA is under control of the HOX proteins. Therefore, one of the regulatory pathways necessary for programmed morphogenesis would be described hierarchically as follows : expression of the homeobox genes-- control of the DNA replication genes-- regulation of the cell proliferation-- normal development.
On the assamption that variable number of tandem repeat(VNTR)sequence may encode hotspots for recombinational activity and be involved in the genesis of polygene abnormality, we used VNTR marker(pYNH24)for the screening of polygene abnormality in congenital malformations. The PYNH24 probe successfully detected abnormal bands in the DNA sample from case MY.
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