1994 Fiscal Year Final Research Report Summary
Structure and expression of the human 2-oxoglutarate dehydrogenase gene
| Project/Area Number |
05670149
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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 |
Pathological medical chemistry
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| Research Institution | Nagasaki University |
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Principal Investigator |
KOIKE Kichiko Nagasaki University Sch.of Med.Associ.Prof., 医学部, 助教授 (80039619)
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| Co-Investigator(Kenkyū-buntansha) |
MATSUO Sachiko Nagasaki University, Inst.Trop.Med.Instructor, 熱帯医学研究所, 助手 (80039916)
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| Project Period (FY) |
1993 – 1994
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| Keywords | 2-oxoglutarate dehydrogenase / gene / exon / intron / coenzyme TPP / transcription start point / chromosomal localization |
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| Research Abstract |
The 2-oxoglutarate dehydrogenase (OGDH) complex that catalyzes oxidative decarboxylation of 2-oxoglutarate. It is apparent that the OGDH complex is composed of three different component enzymes. The OGDH,one of the component enzymes of the complex catalyzes the TPP-dependent irreversible decarboxylation and the subsequent reductive succinylation of the lipoyl moiety. We recentry cloned the human cDNA encoding OGDH from fetal liver cDNA library. To obtain further insights into the regulation of its expression, we tried to isolate genomic OGDH clones using the human cDNA as a probe and characterize the OGDH gene structure.
Genomic clones covering the entire sequence of the gene encoding human OGDH were isolated by screening leukocyte and placenta genomic libraries with radio- and digoxigenin-labeled human OGDH cDNAs. The human OGDH gene contains 22 exons spanning approximately 85 kb. All exon/intron splice junctions follow the GT/AG rule. The results of primer extension analysis imply that the OGDH gene transcription start point is a thymine base 55 bp upstream of the ATG start codon. The 5'-flanking region of the OGDH gene lacked canonical TATA or CAAT boxes. Using DNAs from human * rodent somatic cell hybrids that segregate human chromosome in conjunction with fluorescence in situ hybridization, human OGDH gene was assigned to chromosome arm 7p, an area that corresponds to the boundary between bands 7p13 and 7p14.
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