Molecular cloning of human and mouse DJ-1 genes and identification of Sp1-dependent activation of the human DJ-1 promoter.
DJ-1 has been identified as a novel oncogene that transforms mouse NIH3T3 cells in cooperation with activated ras. Subsequently, two other groups have identified SP22 or CAP-1, rat homologs of human DJ-1, as a sperm protein targeted by some toxicants leading to male infertility, and another group has also reported that RS, the same as human DJ-1, is a component of an RNA-binding protein complex. To characterize the expression or functional importance of DJ-1, the genomic DNAs of both human and mouse DJ-1 were cloned and characterized. Both genomic DNAs comprise 7 exons spanning about 16-24 kb, in which 2-6 exons encode the DJ-1 protein. The human DJ-1 gene was mapped at chromosome 1p36.2-p36.3, a region that has been shown to be a hot spot of chromosome abnormalities in several tumor cells. To analyze the promoter of the human DJ-1 gene, a series of deletion constructs of the region upstream of exon 2 were linked to the luciferase gene, and their luciferase activities were measured in human HeLa cells. Of the many putative transcription regulatory sequences, the Sp1 site present at -100 from the transcription initiation site contributed to the major promoter activity, and Sp1 was identified as a protein binding to this site by a mobility shift assay using HeLa nuclear extract.
Taira
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Graduate School of Pharmaceutical Sciences, Hokkaido University, Kita 12, Nishi 6, Kita-ku, Sapporo 060-0812, Japan.
Takahashi
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Kitagawa
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Iguchi-Ariga
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SM
Ariga
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002874:Chromosome Mapping; D002878:Chromosomes, Human, Pair 1 / Q000235:genetics; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D005091:Exons; D005796:Genes / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D007438:Introns; D020014:K562 Cells; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D015513:Oncogene Proteins / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D000071617:Protein Deglycase DJ-1; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D016329:Sp1 Transcription Factor / Q000378:metabolism; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured; D020298:U937 Cells
D018076:DNA, Complementary; D047908:Intracellular Signaling Peptides and Proteins; D015513:Oncogene Proteins; D011993:Recombinant Fusion Proteins; D016329:Sp1 Transcription Factor; D004247:DNA; D008156:Luciferases; C105131:PARK7 protein, human; D000071617:Protein Deglycase DJ-1