TY - JOUR
T1 - Ku80 gene expression is Sp1-dependent and sensitive to CpG methylation within a novel cis element
AU - Ludwig, Dale L.
AU - Chen, Fanqing
AU - Peterson, Scott R.
AU - Nussenzweig, Andre
AU - Li, Gloria C.
AU - Chen, David J.
N1 - Funding Information:
This work was supported by the US Department of Energy and by N.I.H. grant CA50519 to D.J.C and N.I.H. grants CA31397 and CA56909 to G.C.L.
PY - 1997/10/15
Y1 - 1997/10/15
N2 - The Ku70/80 complex, known as Ku, constitutes the DNA end binding component of the DNA-dependent protein kinase (DNA-PK). We have characterized the promoter region of the mouse and human Ku80 genes to delineate transcriptional elements necessary for basal gene expression and proliferation-dependent regulation. Consensus Sp1 recognition elements were identified in both promoters, and were determined to be essential for basal expression. We further identified a near-perfect palindrome of 21 base pairs located immediately 5' to one Sp1 element. This sequence was present once within the mouse Ku80 promoter and seven times, in a head-to-tail tandem array, within the human Ku80 promoter. This sequence possessed homology with a methylationsensitive promoter element, Enh2, present in the LTR of mouse intractisternal A-particles. Promoter deletion studies and expression analysis of in-vitro methylated reporter gene constructs provided strong evidence that, in vivo, this repeat sequence regulates Ku80 gene expression in cis, through a mechanism involving CpG methylation. Evidence is also presented, suggesting that Ku is directly involved in this regulatory process.
AB - The Ku70/80 complex, known as Ku, constitutes the DNA end binding component of the DNA-dependent protein kinase (DNA-PK). We have characterized the promoter region of the mouse and human Ku80 genes to delineate transcriptional elements necessary for basal gene expression and proliferation-dependent regulation. Consensus Sp1 recognition elements were identified in both promoters, and were determined to be essential for basal expression. We further identified a near-perfect palindrome of 21 base pairs located immediately 5' to one Sp1 element. This sequence was present once within the mouse Ku80 promoter and seven times, in a head-to-tail tandem array, within the human Ku80 promoter. This sequence possessed homology with a methylationsensitive promoter element, Enh2, present in the LTR of mouse intractisternal A-particles. Promoter deletion studies and expression analysis of in-vitro methylated reporter gene constructs provided strong evidence that, in vivo, this repeat sequence regulates Ku80 gene expression in cis, through a mechanism involving CpG methylation. Evidence is also presented, suggesting that Ku is directly involved in this regulatory process.
KW - DNA-dependent protein kinase
KW - Dna repair
KW - Transcription
KW - V(D)J recombination
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U2 - 10.1016/S0378-1119(97)00366-1
DO - 10.1016/S0378-1119(97)00366-1
M3 - Article
C2 - 9358055
AN - SCOPUS:0030817722
SN - 0378-1119
VL - 199
SP - 181
EP - 194
JO - Gene
JF - Gene
IS - 1-2
ER -