Wakabayashi, S. et al

Volume 16 Number 8 1988

Nucleic Acids Research

Nucleotide sequence of a cDNA encoding the precursor to human cytochrome cl Morimitsu Nishikimi, Shigeo Ohtal, Hiroshi Suzuki, Tomoko Tanaka, Fumitaka Kikkawa, Masashi Tanaka, Yasuo Kagawal and Takayuki Ozawa Department of Biomedical Chemistry, Faculty of Medicine, University of Nagoya, Nagoya 466 and 'Department of Biochemistry, Jichi Medical School, Minamikawachi, Tochigi 329-04, Japan Submitted March 16, 1988 Accession no.X06994

A partial nucleotide sequence of a cDNA encoding human cytochrome Ci was previously published (1). Here, we report its complete coding sequence for the precursor to this cytochrome. A human fibroblast (SV-40 transformed cell line, GM637) cDNA library constructed in pcD was screened by colony hybridization, and a positive clone containing a 1.2-kbp cDNA was obtained. This cDNA contains an open reading frame coding for 325 amino acids of the protein with a molecular weight of 35,375. Based on the amino acid sequence of the mature form of bovine cytochrome cl (2), the protein is presumed to have the presequence of 84 amino acids, which are rich in basic amino acids and serine. AGGCCAAG ATG GCG GCA GCT GCG GCT TCG CTT CGC GGG GTA GTG TTG GGC CCG CGG GGC GCG GGG CTC CCG GGC GCG CGT GCC CGG GGT CTG CTG M A A A A A S L R G V V L G P R G A G L P G A R A R G L L AGC GCG CGG CCC GGG CAG CTC CCG CTA CGG ACA CCT CAG GCA GTG GCC TTG TCG TCG AAG TCT GGC CTT TCC CGA GGC CGG AAA GTG S A R P G Q L P L R T P Q A V A L S S K S G L S R G R K V CTG TCA GCG CTG GGC ATG CTG GCG GCA GGG GGT GCG GGG CTG GCC GTG GCT CTG CAT TCG GCT GTG AGT GCC AGT GAC CTG GAG GTG L S A L G M L A A G G A G L A V A L H S A V S A S D L E V CCC CCC AGC TAT CCG TGG TCT CAC CGT GGC CTC CTC TCT TCC TTG GAC CAC ACC AGC ATC CGG AGG GGT TTC CAG GTA TAT AAG CAG PP S Y P W S H R G L L S S L D H T S I R R G F Q V Y K Q TGC GCC TCC TGC CAC AGC ATG GAC TTC GTG GCC TAC CGC CAC CTG GTG GGC GTG TGC TAC ACG GAG GAT GAA GCT MG GAG CTG GCT C A S C H S M D F V A Y R H L V G V C Y T E D E A K E L A GAG GTG GAG GTT CAA GAC GGC CCC MT GAA GAT GGG GAG ATG TTC ATG CGG CCA GGG AAG CTG TTC GAC TAT TTC CCA AAA CCA TAC E V E V Q D G P N E D G E M F M R P G K L F D Y F P K P Y

TGC C ATG M CAC H GTG V GCG A CCC P

AAC AGT GAG GCT GCT CGA GCT GCC MC AAC GGA GCA TTG CCC CCT GAC CTC AGC TAC ATC GTG CGA GCT AGG CAT GGT GGT GAG GAC TAC N S E A A R A A N N G A L P P D L S Y I V R A R H G G E D Y

GTC TTC TCC CTG CTC ACG GGC TAC TGC GAG CCA CCC ACC GGG GTG TCA CTG CGG GAA GGT CTC TAC TTC AAC CCC TAC TTT CCT GGC CAG V F S L L T G Y C E P P T G V S L R E G L Y F N P Y F P G Q

GCC ATT GCC ATG GCC CCT CCC ATC TAC ACA GAT GTC TTA GAG TTT GAC GAT GGC ACC CCA GCT ACC ATG TCC CAG ATA GCC MG GAT GTG A I A M A P P I Y T D V L E F D D G T P A T M S Q I A K D V TGC ACC TTC CTG CGC TGG GCA TCT GAG CCA GAG CAC GAC CAT CGA AAA CGC ATG GGG CTC MG ATG TTG ATG ATG ATG GCT CTG CTG GTG C T F L R W A S E P E H D H R K R M G L K M L M M M A L L V

CCC CTG GTC TAC ACC ATA AAG CGG CAC MG TGG TCA GTC CTG MG AGT CGG MG CTG GCA TAT CGG CCG CCC AAG TGA CCCTGTCCAGTGTCT P L V Y T I K R H K W S V L K S R K L A Y R P P K

GCTTGCCATCCTGCCAGAACAGGCCCTCAAGCCCAAGAGCCATCCCAGGCCTGTTCAGGCTCAGCTAAGCCTCTCTTCATCTGGAAGAAGAGGCAAGGGGGCAGGAGACCAGGCTCTAG CTCTGGGCCCTCCTTCAGCCCCCATCATGGGAATATTAATTTTCTCMTGT Arrow indicates the start of the mature protein.

Acknowledgments The authors thank Dr. Hiroto Okayama for a human fibroblast cDNA This work was supported in part by Grant-in-Aid No. library. 62617002 for Scientific Research on Priority Areas from the Ministry of Education, Science, and Culture of Japan. References (1) Nishikimi, M. et al. (1987) Biochem. Biophys. Res. Commun. 145, 34-39. (2) Wakabayashi, S. et al. (1980) Biochem. Biophys. Res. Commun. 97, 1548-1554. © R L Press Limited, Oxford, England.

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