Structure and Expression of a Complementary DNA for the Nuclear Coded Precursor of Human Mitochondrial Ornithine Transcarbamylase

American Association for the Advancement of Science (AAAS) - Tập 224 Số 4653 - Trang 1068-1074 - 1984
Arthur L. Horwich1, Wayne A. Fenton1, Kenneth R. Williams2, František Kalousek1, Jan P. Kraus1, Russell F. Doolittle3, William H. Konigsberg2, León E. Rosenberg1
1Department of Human Genetics, Yale University School of Medicine, New Haven, Connecticut 06510.
2Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, Connecticut 06510.
3Department of Chemistry, University of California, San Diego, La Jolla, California 92093

Tóm tắt

Most mitochondrial proteins are encoded in the nucleus and are translated on free cytoplasmic ribosomes as larger precursors containing amino-terminal "leader" sequences, which are removed after the precursors are taken up by mitochondria. We have deduced the complete primary structure of the precursor of a human mitochondrial matrix enzyme, ornithine transcarbamylase (OTC), from the nucleotide sequence of cloned complementary DNA. The amino-terminal leader peptide of OTC is 32 amino acids in length and contains four arginines but no acidic residues. Cleavage of the leader peptide from the "mature" protein occurs between glutamine and asparagine residues. The sequence of mature human OTC resembles that of the subunits of both OTC and aspartate transcarbamylase from Escherichia coli. The biological activity of the cloned OTC complementary DNA was tested by joining it with SV40 (an animal virus) regulatory elements and transfecting cultured HeLa cells, which do not normally express OTC. Both the precursor and mature forms of the OTC subunit were identified; in stable transformants, enzymatic activity was also detected.

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American Association for the Advancement of Science (AAAS)

Tập 224 Số 4653

1068-1074

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