Structural organization of the mouse DNA repair gene, N-methylpurine-DNA glycosylase.
N-Methylpurine-DNA glycosylase (MPG), a ubiquitous DNA repair enzyme, removes N-alkylpurines and other purine lesions induced in DNA by simple alkylating carcinogens. A mouse MPG cDNA clone was isolated from a lambda recombinant phage library of BALB/c mouse lung cell and characterized. Using the mouse MPG cDNA as a probe, the complete mouse MPG gene was isolated in two overlapping lambda recombinant genomic clones. The 6-kb gene has four exons containing 1,002 bp of coding sequence. The transcription start site was identified in the genomic sequence by primer extension of MPG mRNA from a mouse lung fibroblast cell line. The location of this transcription start site was confirmed by in vitro transcription with the promoter-containing plasmid template. Promoter function of the sequence 5' upstream of the transcription initiation site was shown by transient expression of the firefly luciferase reporter gene under the control of this sequence in transfected human and mouse cells. The mouse MPG promoter contains no TATA box, but has a CAAT element and is G.C-rich with putative AP2 elements and SP1-complementary sequences.
Tatsuka
M
M
Sealy Center for Molecular Science, University of Texas Medical Branch, Galveston 77555.
Ibeanu
G C
GC
Izumi
T
T
Narayan
S
S
Ramana
C V
CV
Kim
N K
NK
Kang
W
W
Roy
G
G
Mitra
S
S
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D045647:DNA Glycosylases; D004260:DNA Repair / Q000235:genetics*; D005091:Exons / Q000235:genetics; D015723:Gene Library; D008168:Lung; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009699:N-Glycosyl Hydrolases / Q000737:chemistry / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic / Q000235:genetics
D012333:RNA, Messenger; D045647:DNA Glycosylases; D009699:N-Glycosyl Hydrolases; C076131:DNA-3-methyladenine glycosidase II