An initiator element and a proximal cis-acting sequence are essential for transcriptional activation of the complement factor I (CFI) gene.
Human complement factor I (CFI) is a serine protease which regulates the complement system by inactivation of C3b and C4b in the presence of appropriate cofactors. In this study, we have analyzed the mechanism controlling the constitutive transcriptional activation of the CFI gene. Using deletion analysis and transient CAT expression assays, we have mapped the minimal promoter to the region located between -46 and +160 bp relative to the major transcription start point (tsp), and also shown that cis-acting elements both upstream and downstream of the tsp are important for promoter activity. A silencer element was also found between -71 and -46 bp. The sequence surrounding the tsp was related to the mouse terminal deoxynucleotidyltransferase initiator element (Inr) and point mutations in this sequence, from -3 to +4, drastically reduced CFI promoter activity. Mutations in a -9 to -5 bp CTGAA sequence immediately upstream of the tsp also reduced promoter activity. Gel mobility shift analysis demonstrated the binding of nuclear factors to a CTGAA repeat located at -9 to -5 and +101 to +105. Our results suggest that CFI promoter contains a functional Inr element that is essential for promoter activity, and the interactions of the CTGAA element located between -9 and +5 with nuclear factor(s) may be part of the machinery required for CFI Inr-dependent transcription.
Paramaswara
B
B
Department of Laboratory Medicine and Pathobiology, University of Toronto, Canada.
Minta
J O
JO
D000818:Animals; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular / Q000235:genetics; D002457:Cell Extracts; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D017244:Complement Factor I / Q000235:genetics* / Q000378:metabolism; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004253:DNA Nucleotidylexotransferase / Q000235:genetics; D006801:Humans; D051379:Mice; D016297:Mutagenesis, Site-Directed; D009841:Oligonucleotides / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid; D017384:Sequence Deletion; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic; D015533:Transcriptional Activation / Q000235:genetics*; D014162:Transfection
D002457:Cell Extracts; D009841:Oligonucleotides; D011994:Recombinant Proteins; D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase; D004253:DNA Nucleotidylexotransferase; D017244:Complement Factor I