ISOLATION, MAPPING, AND GENOMIC STRUCTURE OF AN X-LINKED GENE FOR A SUBUNIT OF HUMAN MITOCHONDRIAL COMPLEX-I

We report here the isolation, mapping, and genomic organization of the human NDUFA1 gene, which is a component of NADH:ubiquinone oxidoreduc tase (complex I). The NDUFA1 cDNA clone and associated genomic cosmid clones were isolated by reciprocal probing of an arrayed human heart c DNA Library with a X-chromosome cosmid library and were mapped to Xq24 . The NDUFA1 gene, which is highly expressed in human cardiac and skel etal muscle, has an open reading frame of 70 amino acids and shows 80% homology to the bovine MWFE subunit of complex I. By primer extension , the major and minor transcription initiation sites were identified, 99 and 141 nucleotides upstream of the translation initiator ATG, resp ectively. The NDUFA1 gene is composed of 3 exons and spans about 5.0 k b of genomic DNA. The 5' region of the NDUFA1 gene (similar to 450-bp fragment) lacks conventional TATA and CAAT boxes, but it contains seve ral potential binding sites for transcription factors including SP-1, AP-2, NF1, NRF2-like, APRRE, CRE, MyoD1, CArG, MEF-2, and BRE. (C) 199 6 Academic Press, Inc.