P. Jackson et al., SYNERGISTIC TRANSCRIPTIONAL ACTIVATION OF THE MCK PROMOTER BY P53 - TETRAMERS LINK SEPARATED DNA RESPONSE ELEMENTS BY DNA LOOPING, Oncogene, 16(2), 1998, pp. 283-292
The WAF1, Cyclin G and muscle creatine kinase (MCK) genes, all contain
multiple copies of the consensus p53-binding element within their reg
ulatory regions, We examined the role of these elements in transactiva
tion of the muscle creatine kinase (MCK) gene by p53, The MCK promoter
possesses distal (-3182 to -3133) and proximal (-177 to -81) p53-bind
ing elements within which residues -3182 to -3151 (distal) and -176 to
-149 (proximal) show homology to the consensus p53-binding site, Usin
g promoter deletion studies, we find that both proximal and distal ele
ments are required for high level, synergistic transcriptional activat
ion in vivo. Electron microscopy indicates that p53-p53 interactions l
ink proximal and distal p53-binding elements and cause looping out of
intervening DNA, suggesting that this DNA sequence may be dispensable
for synergy, This idea was confirmed by progressive deletion of the DN
A between p53-binding elements, Synergism persisted with spacing reduc
ed to only 150 bp, Tetramerization-deficient p53 mutants were defectiv
e for transcriptional activation but still capable of synergy, Our res
ults provide evidence for a model by which high level transcriptional
activation of promoters with multiple p53 response elements is achieve
d.