TY - JOUR
T1 - Positive and negative regulation of transcription in vitro
T2 - Enhancer-binding protein AP-2 is inhibited by SV40 T antigen
AU - Mitchell, Pamela J.
AU - Wang, Charlotte
AU - Tjian, Robert
N1 - Funding Information:
We thank William Dynan for kindly providing SV40 linker-scanning plasmid constructions LS11, LS12, and LS13 prior to publication; Pierre Chambon for SV40 plasmids pA0, pA12, and pA15; and Winship Herr for SV40 mutant constructions dpm8 and dpml2, which allowed our initial identification of factor AP-3. Stimulating discussions, technical advice, and critical readings of this manuscript provided by Jim Ka-donaga, Winship Herr, Dirk Bohmann, and other members of the Tjian lab contributed significantly to this work and were greatly appreciated. R. Tjian is partly funded by a research grant from the National Cancer Institute, and P J. Mitchell is a Damon Runyon-Walter Winchell Fellowship recipient.
PY - 1987/9/11
Y1 - 1987/9/11
N2 - We have purified a 52 kd protein, AP-2, that binds to enhancer regions of SV40 and human metallothionein IIA (hMT IIA) and stimulates RNA synthesis from these promoters in vitro. Surprisingly, AP-2 also binds to two SV40 early promoter regions recognized by Sp1 and T antigen. Juxtaposed binding sites for AP-2 and Sp1 in the 21 bp repeats may facilitate productive interactions between the two factors. In contrast, sequence-specific binding of AP-2 to SV40 and hMT IIA DNA is inhibited by the viral repressor protein T antigen. Furthermore, T antigen inhibits AP-2-dependent transcriptional activation of the hMT IIA promoter in vitro. The inhibition is neither a direct nor an indirect result of T antigen binding to DNA, because the hMT IIA promoter lacks T antigen binding sites. Instead, sedimentation studies suggest that protein-protein interactions between AP-2 and T antigen block AP-2 binding to DNA. These findings suggest novel mechanisms for mediating positive and negative regulation of transcription.
AB - We have purified a 52 kd protein, AP-2, that binds to enhancer regions of SV40 and human metallothionein IIA (hMT IIA) and stimulates RNA synthesis from these promoters in vitro. Surprisingly, AP-2 also binds to two SV40 early promoter regions recognized by Sp1 and T antigen. Juxtaposed binding sites for AP-2 and Sp1 in the 21 bp repeats may facilitate productive interactions between the two factors. In contrast, sequence-specific binding of AP-2 to SV40 and hMT IIA DNA is inhibited by the viral repressor protein T antigen. Furthermore, T antigen inhibits AP-2-dependent transcriptional activation of the hMT IIA promoter in vitro. The inhibition is neither a direct nor an indirect result of T antigen binding to DNA, because the hMT IIA promoter lacks T antigen binding sites. Instead, sedimentation studies suggest that protein-protein interactions between AP-2 and T antigen block AP-2 binding to DNA. These findings suggest novel mechanisms for mediating positive and negative regulation of transcription.
UR - http://www.scopus.com/inward/record.url?scp=0023651331&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0023651331&partnerID=8YFLogxK
U2 - 10.1016/0092-8674(87)90512-5
DO - 10.1016/0092-8674(87)90512-5
M3 - Article
C2 - 3040262
AN - SCOPUS:0023651331
SN - 0092-8674
VL - 50
SP - 847
EP - 861
JO - Cell
JF - Cell
IS - 6
ER -