TY - JOUR
T1 - Molecular and functional analysis of the XPBC/ERCC-3 promoter
T2 - Transcription activity is dependent on the integrity of an SP1-binding site
AU - Ma, Libin
AU - Weeda, Geert
AU - Jochemsen, Aart G.
AU - Bootsma, Dirk
AU - Hoeijmakers, Jan H.J.
AU - van der Eb, Alex J.
N1 - Funding Information:
We would like to thank H. van Ormondt for his critical reading of the manuscript. We are grateful to H. van Dam for his help in band shift assays and I. A. Laird-Offringa for her help in RNA analysis. We also thank H. van Dam, LA. Laird-Offringa, S.J.L. van den Heuvel, and A. Zantema for their helpful discussions. This work was supported by the J.A. Cohen Institute for Radiopathology and Radiation Protection (IRS).
PY - 1992/1/25
Y1 - 1992/1/25
N2 - The human XPBC/ERCC-3 gene, which corrects the excision-repair defect in xeroderma pigmentosum group B cells and the UV-sensitive CHO mutant 27-1 cells, appears to be expressed constitutively in various cell types and tissues. We have analysed the structure and functionality of the XPBC/ERCC-3 promoter. Transcription of the XPBC/ERCC-3 gene is initiated from heterogeneous sites, with a major startpoint mapped at position - 54 (relative to the translation start codon ATG). The promoter region does not possess classical TATA and CAAT elements, but it is GC-rich and contains three putative Sp1-binding sites. In addition, there are two elements related to the cyclic AMP (cAMP)-response element (CRE) and the 12-O-tetradecanoyl phorbol-13-acetate-response element (TRE) in the 5'-fIanking region. Transient expression analysis of XPBC/ERCC-3 promoter-CAT chimeric plasmids revealed that a 127-bp fragment, spanning position -129 to -3, is minimally required for the promoter activity. Transcription of the XPBC/ERCC-3 promoter depends on the integrity of a putative Sp1-binding site in close proximity to the major cap site. Band shift assays showed that this putative Sp1-binding site can interact specifically with a nuclear factor, most likely transcription factor Sp1 (or an Sp1-like factor) in vitro.
AB - The human XPBC/ERCC-3 gene, which corrects the excision-repair defect in xeroderma pigmentosum group B cells and the UV-sensitive CHO mutant 27-1 cells, appears to be expressed constitutively in various cell types and tissues. We have analysed the structure and functionality of the XPBC/ERCC-3 promoter. Transcription of the XPBC/ERCC-3 gene is initiated from heterogeneous sites, with a major startpoint mapped at position - 54 (relative to the translation start codon ATG). The promoter region does not possess classical TATA and CAAT elements, but it is GC-rich and contains three putative Sp1-binding sites. In addition, there are two elements related to the cyclic AMP (cAMP)-response element (CRE) and the 12-O-tetradecanoyl phorbol-13-acetate-response element (TRE) in the 5'-fIanking region. Transient expression analysis of XPBC/ERCC-3 promoter-CAT chimeric plasmids revealed that a 127-bp fragment, spanning position -129 to -3, is minimally required for the promoter activity. Transcription of the XPBC/ERCC-3 promoter depends on the integrity of a putative Sp1-binding site in close proximity to the major cap site. Band shift assays showed that this putative Sp1-binding site can interact specifically with a nuclear factor, most likely transcription factor Sp1 (or an Sp1-like factor) in vitro.
UR - http://www.scopus.com/inward/record.url?scp=0026575426&partnerID=8YFLogxK
U2 - 10.1093/nar/20.2.217
DO - 10.1093/nar/20.2.217
M3 - Article
C2 - 1741247
AN - SCOPUS:0026575426
SN - 0305-1048
VL - 20
SP - 217
EP - 224
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 2
ER -