PMID: 9173885Apr 1, 1997Paper

Transcriptional activation of the minimal human Proalpha1(I) collagen promoter: obligatory requirement for Sp1

The Biochemical Journal
H M Poppleton, R Raghow

Abstract

A construct containing human Proalpha1(I) collagen gene promoter/enhancer-driven chloramphenicol acetyltransferase (CAT), pCOL-KT, failed to be expressed significantly in Sp1-deficient Schneider Drosophila line 2 (SL2) cells. However, CAT expression was induced 200-fold in SL2 cells co-transfected with pCOL-KT and pPACSp1, an Sp1-expression vector driven by the Drosophila actin 5C promoter. Elimination of the four potential Sp1-binding sites from pCOL-KT (pCOL-KTDeltaI), by removal of the first intron, did not abrogate Sp1-mediated induction of CAT. Even more significantly, a minimal Proalpha1(I) collagen promoter (-100 to +117 bp), containing a TATA box (-28 to -25 bp) and one putative Sp1-binding site (-87 to -82 bp), elicited strong Sp1-induced transactivation. Furthermore, mutation of the Sp1 motif in the minimal Proalpha1(I) collagen promoter-CAT construct abolished Sp1-induced expression of the reporter gene. Purified Sp1 protein bound specifically to DNA fragments of the Proalpha1(I) minimal promoter encompassing the putative Sp1-binding site; Sp1 binding could be competed out by a double-stranded oligonucleotide containing the wild-type Sp1 sequence, while an oligonucleotide containing a mutated Sp1 site failed to compe...Continue Reading

Citations

Sep 8, 2000·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·C VermesT T Glant
Mar 30, 2001·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·K A RoebuckT T Glant
Oct 2, 2003·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·C BergeronJ Chakir
Jan 18, 2006·Biochemical and Biophysical Research Communications·Stefan BeckertM Zamirul Hussain

❮ Previous
Next ❯

Related Concepts

Related Feeds

CREs: Gene & Cell Therapy

Gene and cell therapy advances have shown promising outcomes for several diseases. The role of cis-regulatory elements (CREs) is crucial in the design of gene therapy vectors. Here is the latest research on CREs in gene and cell therapy.