The promoter of the C1 inhibitor gene contains a polypurine·polypyrimidine segment that enhances transcriptional activity

Kamyar Zahedi, John Bissler, Anne E. Prada, Jorge A. Prada, Alvin E. Davis

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The C1 inhibitor (C1INH) promoter is unusual in two respects: 1) It contains no TATA sequence, but instead contains a TdT-like initiator element (Inr) at nucleotides -3 to +5; 2) it contains a polypurine·polypyrimidine tract between nucleotides -17 and -45. Disruption of the Inr by the introduction of point mutations reduced promoter activity by 40%. A TATA element inserted at nucleotide -30 in the wild-type promoter and in promoter constructs containing the mutated Inr led to a 2-fold increase in basal promoter activity. Previous studies suggested that the potential hinged DNA- forming polypurine·polypyrimidine tract might be important in the regulation of C1INH promoter activity. The present studies indicate that this region is capable of such intramolecular triple helix formation. Disruption of the polypurine·polypyrimidine sequence by substitution of 5 of the 23 cytosine residues with adenine prevented triple helix formation. Site-directed mutagenesis experiments demonstrate that the regulation of promoter activity is independent of hinged DNA-forming capacity but requires an intact AC box (ACCCTNNNNNACCCT) or the overlapping PuF binding site (GGGTGGG). The C1INH gene also contains a number of potential regulatory elements, including an Sp-1 and an hepatocyte nuclear factor-1 binding site and a CAAT box. The role of these elements in regulation of the C1INH promoter was examined. Elimination of the hepatocyte nuclear factor-1 site at nucleotides -94 to - 81 by truncation reduced the activity of the promoter by ~50%. Similarly, site-directed mutations that disrupt this site reduce promoter activity by 70%.

Original languageEnglish (US)
Pages (from-to)7249-7255
Number of pages7
JournalJournal of Immunology
Volume162
Issue number12
StatePublished - Jun 15 1999
Externally publishedYes

Fingerprint

Hepatocyte Nuclear Factor 1
Nucleotides
Genes
Binding Sites
Cytosine
DNA
Adenine
Site-Directed Mutagenesis
Point Mutation
Mutation

All Science Journal Classification (ASJC) codes

  • Immunology

Cite this

The promoter of the C1 inhibitor gene contains a polypurine·polypyrimidine segment that enhances transcriptional activity. / Zahedi, Kamyar; Bissler, John; Prada, Anne E.; Prada, Jorge A.; Davis, Alvin E.

In: Journal of Immunology, Vol. 162, No. 12, 15.06.1999, p. 7249-7255.

Research output: Contribution to journalArticle

Zahedi, Kamyar ; Bissler, John ; Prada, Anne E. ; Prada, Jorge A. ; Davis, Alvin E. / The promoter of the C1 inhibitor gene contains a polypurine·polypyrimidine segment that enhances transcriptional activity. In: Journal of Immunology. 1999 ; Vol. 162, No. 12. pp. 7249-7255.
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abstract = "The C1 inhibitor (C1INH) promoter is unusual in two respects: 1) It contains no TATA sequence, but instead contains a TdT-like initiator element (Inr) at nucleotides -3 to +5; 2) it contains a polypurine·polypyrimidine tract between nucleotides -17 and -45. Disruption of the Inr by the introduction of point mutations reduced promoter activity by 40{\%}. A TATA element inserted at nucleotide -30 in the wild-type promoter and in promoter constructs containing the mutated Inr led to a 2-fold increase in basal promoter activity. Previous studies suggested that the potential hinged DNA- forming polypurine·polypyrimidine tract might be important in the regulation of C1INH promoter activity. The present studies indicate that this region is capable of such intramolecular triple helix formation. Disruption of the polypurine·polypyrimidine sequence by substitution of 5 of the 23 cytosine residues with adenine prevented triple helix formation. Site-directed mutagenesis experiments demonstrate that the regulation of promoter activity is independent of hinged DNA-forming capacity but requires an intact AC box (ACCCTNNNNNACCCT) or the overlapping PuF binding site (GGGTGGG). The C1INH gene also contains a number of potential regulatory elements, including an Sp-1 and an hepatocyte nuclear factor-1 binding site and a CAAT box. The role of these elements in regulation of the C1INH promoter was examined. Elimination of the hepatocyte nuclear factor-1 site at nucleotides -94 to - 81 by truncation reduced the activity of the promoter by ~50{\%}. Similarly, site-directed mutations that disrupt this site reduce promoter activity by 70{\%}.",
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