Involvement of the transcriptional factor GATA-1 in regulation of expression of coproporphyrinogen oxidase in mouse erythroleukemia cells.
Coproporphyrinogen oxidase (CPO; EC 1.3.3.3), the sixth enzyme of heme biosynthesis, transcribed from a single promoter is markedly induced during erythroid differentiation. CPO is ubiquitously expressed in all cells. To determine cis-acting elements of the human CPO gene, the promoter region of the gene was isolated, and three potential GATA-1 motifs and four GC boxes were found within this fragment. In a functional analysis of various deletion mutants, we found that the GATA-1 binding site at -143 to -138 was essential for basic and inducible expressions of the CPO gene in mouse erythroleukemia (MEL) cells. Gel mobility shift assay revealed that GATA-1 bound to the region is required for the expression and this was confirmed by observations that the nuclear protein bound to the GATA-1 motif was supershifted with anti GATA-1 antibody, by gel mobility shift assay. Furthermore, co-expression of mouse GATA-1 in MEL cells led to an increase in the promoter activity, which was markedly increased by dimethyl sulfoxide-treatment. These results indicate that GATA-1 plays an important role in regulation of transcription of the CPO gene in erythroid cells.
Tanabe
A
A
Department of Hygiene, Kansai Medical University, Osaka, Japan.
Furukawa
T
T
Ogawa
Y
Y
Yamamoto
M
M
Hayashi
N
N
Tokunaga
R
R
Taketani
S
S
D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation / Q000187:drug effects; D003304:Coproporphyrinogen Oxidase / Q000096:biosynthesis / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004273:DNA, Neoplasm / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D050981:Erythroid-Specific DNA-Binding Factors; D004920:Erythropoiesis / Q000235:genetics / Q000502:physiology; D050982:GATA1 Transcription Factor; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000201:enzymology* / Q000235:genetics* / Q000473:pathology; D051379:Mice; D008969:Molecular Sequence Data; D017384:Sequence Deletion; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D018076:DNA, Complementary; D004273:DNA, Neoplasm; D004268:DNA-Binding Proteins; D050981:Erythroid-Specific DNA-Binding Factors; D050982:GATA1 Transcription Factor; C494688:GATA1 protein, human; C494689:Gata1 protein, mouse; D014157:Transcription Factors; D003304:Coproporphyrinogen Oxidase; D004121:Dimethyl Sulfoxide