Isolation of a functional human gene for brain creatine kinase.
There is evidence that the gene for the B isozyme of creatine kinase is regulated during cell differentiation, is under hormonal control, and is activated in a small cell lung carcinoma. In order to investigate further the mechanisms of these processes, the human gene was isolated and the structure of the promoter region was determined. A human DNA fragment of 8 kilobase pairs was shown to encompass the entire coding region and 850 base pairs (bp) of the 5'-flanking sequence. This fragment was transfected into three cell lines and shown to express functional enzyme. The 5'-end of the gene is split by a 230-bp intron that is located 12 bp upstream of the initiator ATG codon. Transcription initiation occurs at a site that is approximately 69 bp upstream of the 5'-end of this intron. The DNA sequence in the region upstream of the 5'-end of the mRNA is suggestive of two superimposed promoters that contain additional sequence elements that are known to regulate expression of other eukaryote genes. The 5'-region also has a remarkable homology to the overlapping promoters of the adenovirus EIIaE gene. These elements collectively form the basis for initial investigations of how this gene is controlled.
Daouk
G H
GH
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
Kaddurah-Daouk
R
R
Putney
S
S
Kingston
R
R
Schimmel
P
P
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000201:enzymology*; D002478:Cells, Cultured; D003402:Creatine Kinase / Q000235:genetics*; D004262:DNA Restriction Enzymes / Q000378:metabolism; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D014162:Transfection
D003402:Creatine Kinase; D004262:DNA Restriction Enzymes