Cloning and functional characterization of the 5'-flanking region of human methionine adenosyltransferase 2A gene.
Methionine adenosyltransferase (MAT) is a critical cellular enzyme which catalyzes the formation of S-adenosylmethionine, the principal methyl donor. In mammals, two different genes, MAT1A and MAT2A, encode for liver-specific and non-liver-specific MAT, respectively. We have cloned and characterized a 1.4-kb 5'-flanking region of the human MAT2A (GenBank Accession No. AF039088). Two major transcriptional start sites were identified by primer extension and S1 nuclease protection analysis; one was within 10 nucleotides downstream and the other was located at 158 nucleotides upstream from the consensus TATA box, respectively. The promoter is highly GC rich (75%) in the first 300 base pairs and contains several Sp-1 binding sites, a C/EBP, a HSF2, a STATx, a c-Myb, several v-Myb, and numerous GATA consensus binding sites. The human MAT2A promoter was able to efficiently drive luciferase expression in both Jurkat and 293 cells, but sequential deletion analysis of the promoter revealed that different regions of the promoter are important for cell-specific MAT2A expression.
Department of Medicine, University of Southern California School of Medicine, Los Angeles 90033, USA.
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D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004268:DNA-Binding Proteins / Q000378:metabolism; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis / Q000235:genetics*; D019169:Jurkat Cells; D008156:Luciferases / Q000096:biosynthesis; D008716:Methionine Adenosyltransferase / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D020598:Proto-Oncogene Proteins c-myb; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D017384:Sequence Deletion; D015534:Trans-Activators / Q000378:metabolism; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D007527:Isoenzymes; D011518:Proto-Oncogene Proteins; D020598:Proto-Oncogene Proteins c-myb; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D008156:Luciferases; D008716:Methionine Adenosyltransferase