Structural analysis of human pyruvate kinase L-gene and identification of the promoter activity in erythroid cells.
The human pyruvate kinase (PK) L-gene is organized in 12 exons over 9.5 kilobases, and the first and second exons are specifically transcribed to the R- and L-type PK mRNA. The 5'-flanking region upstream the first exon has two CAC boxes and four GATA motifs within 250 bp from the translational initiation codon. Comparison with the rat L-gene revealed four well-conserved elements in the region. The transient transfection demonstrated that the 270-bp upstream region was a powerful promoter in K562 cells, whereas deletion of the distal 150-bp sequences, which included three GATA motifs, resulted in drastic reduction of the activity. When the hypersensitive site 2 of the human beta-globin gene locus was joined to the promoter, the activity of the proximal 120-bp region was enhanced. We concluded that the proximal 120-bp region had a basal promoter activity and that the distal 150-bp region acted as an enhancer in erythroid cells.
Kanno
H
H
Okinaka Memorial Institute for Medical Research, Japan.
Fujii
H
H
Miwa
S
S
D000818:Animals; D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D005091:Exons; D005796:Genes; D015698:Genomic Library; D006801:Humans; D007438:Introns; D007527:Isoenzymes / Q000235:genetics*; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011770:Pyruvate Kinase / Q000235:genetics*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D014407:Tumor Cells, Cultured
D007527:Isoenzymes; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; D015500:Chloramphenicol O-Acetyltransferase; D011770:Pyruvate Kinase