Normal transcription of the C1 inhibitor gene is dependent upon a polypurine-polypyrimidine region within the promoter.
Analysis of the transcriptional activity of C1 inhibitor (CIINH) promoter reporter constructs with mutations in the R-Y region indicate that triplex formation by this region is not a predictor of transcriptional activity and that normal promoter function depends on the interaction of trans acting factors with specific elements within this region. Electrophoretic mobility shift assay (EMSA) of Hep3B nuclear extracts using the wild type promoter probe (nucleotides -98 to -9) yielded four major bands. Incubation of the same extracts with probes lacking the HNF-1 site resulted in the disappearance of one band. Supershift assays indicate that HNF-1alpha is the only previously identified protein that is present in the EMSA bands. Southwestern blot analysis detected four bands (M(r)s -130, 75, 65 and 20 kDa). These data suggest that the -98 to -9 region of the C1INH promoter interacts with at least four proteins, one of which is HNF-1alpha.
Division of Nephrology and Hypertension, Children's Hospital Research Foundation, Cincinnati, Ohio 45229, USA. zahek0@chmcc.org
Kamyar
K
Zahedi
Anne E
AE
Prada
Aideen
A
Mulligan
Jorge A
JA
Prada
Alvin E
AE
Davis
D026501:Blotting, Southwestern; D003174:Complement C1 Inactivator Proteins; D050718:Complement C1 Inhibitor Protein; D004268:DNA-Binding Proteins / Q000378:metabolism; D004586:Electrophoresis; D015870:Gene Expression / Q000502:physiology; D051537:Hepatocyte Nuclear Factor 1; D051538:Hepatocyte Nuclear Factor 1-alpha; D051539:Hepatocyte Nuclear Factor 1-beta; D006801:Humans; D009154:Mutation / Q000502:physiology; D009687:Nuclear Proteins; D011401:Promoter Regions, Genetic / Q000235:genetics / Q000502:physiology*; D011687:Purines; D011743:Pyrimidines; D015843:Serpins / Q000235:genetics*; D014157:Transcription Factors / Q000502:physiology; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D003174:Complement C1 Inactivator Proteins; D050718:Complement C1 Inhibitor Protein; D004268:DNA-Binding Proteins; C496487:HNF1A protein, human; C496495:HNF1B protein, human; D051538:Hepatocyte Nuclear Factor 1-alpha; D009687:Nuclear Proteins; D011687:Purines; D011743:Pyrimidines; C469952:SERPING1 protein, human; D015843:Serpins; D014157:Transcription Factors; D051537:Hepatocyte Nuclear Factor 1; D051539:Hepatocyte Nuclear Factor 1-beta; C030986:pyrimidine; C030985:purine