Cloning, structure, chromosomal localization and promoter analysis of human 2-oxoglutarate dehydrogenase gene

Abstract

Human 2-oxoglutarate dehydrogenase (OGDH) is an E1-component of the OGDH multi-enzyme complex and catalyzes both the ThDP-dependent decarboxylation of 2-oxoglutarate and the subsequent reductive succinylation of the lipoyl moiety which is covalently bound to the E2 component, dihydrolipoamide succinyltransferase. The cDNA and genomic DNA encoding human OGDH has been cloned and sequenced. The cDNA contains a 3006-bp open reading frame encoding a 40-amino acid leader peptide and a 962-amino acid mature OGDH protein (Mr=108878). The gene contains 22 exons spanning approximately 85 kb. The putative ThDP-binding sequence motif is identified in both DNAs. The gene is localized to chromosome 7 at p13-p14 by fluorescence in situ hybridization. With the TATA- and CAAT-less 5'-flanking region (wild type, -3276/+212) of the OGDH gene-luciferase reporter vector construct and its nested deletion or linker-scanning mutant constructs the transient reporter expression assays in BHK-21 cells reveal the existence of two 10-bp cis-acting elements (-53/-44 and -33/-24) and two trans-acting elements (-536/-496 and -93/-84). A nuclear factor that binds to the region from -63 to -24 including two cis-acting elements.