Binding of f-PIP, a pyrrole- and imidazole-containing triamide, to the inverted CCAAT box-2 of the topoisomerase IIalpha promoter and modulation of gene expression in cells.
An N-formamido pyrrole- and imidazole-containing triamide (f-PIP) has been shown by DNase I footprinting, SPR, and CD studies to bind as a stacked dimer to its cognate sequences: 5'-TACGAT-3' (5'-flank of the inverted CCAAT box-2 of the human topoisomerase IIalpha promoter) and 5'-ATCGAT-3'. A gel shift experiment provided evidence for f-PIP to inhibit protein-DNA interaction at the ICB2 site. Western blot studies showed that expression of the topoisomerase IIalpha gene in confluent NIH 3T3 cells was induced by treatment with f-PIP. The results suggested that the triamide was able to enter the nucleus, interacted with the target site within ICB2, inhibited NF-Y binding, and activated gene expression.
Le
N Minh
NM
Department of Chemistry, Furman University, 3300 Pointsett Highway, Greenville, SC 29613, USA.
Sielaff
Alan M
AM
Cooper
Amanda J
AJ
Mackay
Hilary
H
Brown
Toni
T
Kotecha
Minal
M
O'Hare
Caroline
C
Hochhauser
Daniel
D
Lee
Moses
M
Hartley
John A
JA
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D002942:Circular Dichroism; D018983:DNA Footprinting; D004250:DNA Topoisomerases, Type II / Q000235:genetics*; D003850:Deoxyribonuclease I; D004214:Distamycins / Q000493:pharmacokinetics*; D015972:Gene Expression Regulation, Neoplastic; D007093:Imidazoles / Q000493:pharmacokinetics*; D051379:Mice; D011401:Promoter Regions, Genetic; D011758:Pyrroles / Q000493:pharmacokinetics*
D004214:Distamycins; D007093:Imidazoles; C516205:N-(2-(diethylamino)ethyl)-1-methyl-4-(1-methyl-4-(4-formamido-1-methylimidazole-2-carboxamido)pyrrole-2-carboxamido)imidazole-2-carboxamide; D011758:Pyrroles; D003850:Deoxyribonuclease I; D004250:DNA Topoisomerases, Type II