Cloning and expression of murine S-adenosylmethionine synthetase.
Mice homozygous for chromosomal deletions at or around the albino locus on chromosome 7 express reduced levels of a group of liver genes. Here, we report the isolation and characterization of cDNA and genomic clones encoding one of the affected genes, the mouse adult liver S-adenosylmethionine (AdoMet) synthetase. This enzyme catalyzes the synthesis of AdoMet, which functions in transmethylation and transsulfuration. Mouse AdoMet synthetase cDNA is 3232 base pairs (bp) in length and contains an open reading frame that encodes an enzymatically active polypeptide of 396 amino acids. The mouse AdoMet synthetase shares 98 and 96% amino acid sequence identity with the adult liver enzyme in the rat and human, respectively. AdoMet synthetases possess the consensus ATP-binding motif Gly-X-Gly-X-X-Gly and a putative ATP-binding Lys residue at conserved locations. As an initial step toward understanding the control of AdoMet synthetase gene expression, we characterized the complete transcription unit of this gene. The AdoMet synthetase gene spans approximately 18 kilobases and consists of nine exons ranging from 78 to 1920 bp. The transcription initiation site was demonstrated by rapid amplification of cDNA ends and confirmed by primer extension studies. A putative TATA box is located at -28 to -23 bp upstream of the transcription start site. The cis-acting DNA elements in the 5'-flanking region of the AdoMet synthetase gene that drive chloramphenicol acetyltransferase gene expression in mouse hepatocytes were identified by transient expression assays. The -365 to -2-bp DNA region upstream of the transcription start site of the AdoMet synthetase gene contains promoter elements, and the -518 to -366-bp DNA region might be involved in negative gene regulation.
Sakata
S F
SF
Human Genetics Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892.
Shelly
L L
LL
Ruppert
S
S
Schutz
G
G
Chou
J Y
JY
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D004247:DNA; D005091:Exons; D015870:Gene Expression; D015698:Genomic Library; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008716:Methionine Adenosyltransferase / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000378:metabolism; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid; D014162:Transfection
D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase; D008716:Methionine Adenosyltransferase