Regulated expression of the overlapping ubiquitous and erythroid transcription units of the human porphobilinogen deaminase (PBG-D) gene introduced into non-erythroid and erythroid cells.
The human gene coding for porphobilinogen deaminase (PBG-D) is transcribed into two distinct transcription units giving two mRNAs. These units originate from two adjacent promoters distant of 3 kilobase pairs. The upstream promoter is active in all cell types, whereas the downstream promoter is active only in erythroid cells. We have studied the expression of this gene either after introduction of the corresponding human chromosome into murine erythroid cells using somatic hybrids or after transfection into both erythroid and non-erythroid cells. Using somatic hybrids, we showed that activation of the erythroid-specific promoter of the PBG-D gene did not reduce the rate of initiation of the ubiquitous promoter. Transfection experiments in erythroid cells showed that the PBG-D erythroid transcription unit, controlled by the PBG-D erythroid promoter, was correctly transcribed and regulated. Furthermore, we found that the PBG-D erythroid promoter alone was sufficient for correct expression and regulation of a reporter gene during erythroid differentiation. When the human PBG-D gene was transfected into non-erythroid cells, only the ubiquitous promoter was active. Deletion of the ubiquitous promoter did not lead to any activation of the erythroid promoter, suggesting that its inactivity in non-erythroid cells was not due to promoter occlusion but to a strict erythroid specificity.
Raich
N
N
Institut National de la Santé et de la Recherche Médicale Unité, Hôpital Henri Mondor, Créteil, France.
Mignotte
V
V
Dubart
A
A
Beaupain
D
D
Leboulch
P
P
Romana
M
M
Chabret
C
C
Charnay
P
P
Papayannopoulou
T
T
Goossens
M
M
D000642:Ammonia-Lyases / Q000235:genetics*; D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D004742:Enhancer Elements, Genetic; D005796:Genes; D005914:Globins / Q000235:genetics; D006801:Humans; D011163:Hydroxymethylbilane Synthase / Q000235:genetics*; D004915:Leukemia, Erythroblastic, Acute / Q000201:enzymology; D051379:Mice; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D015183:Restriction Mapping; D013539:Simian virus 40 / Q000235:genetics; D014158:Transcription, Genetic; D014162:Transfection
D012333:RNA, Messenger; D005914:Globins; D011163:Hydroxymethylbilane Synthase; D000642:Ammonia-Lyases