In vivo mapping of the human adenine nucleotide translocator-2 (ANT2) promoter provides support for regulation by a pair of proximal Sp1-activating sites and an upstream silencer element

被引:8
作者
Luciakova, K
Hodny, Z
Barath, P
Nelson, BD [1 ]
机构
[1] Univ Stockholm, Arrhenius Lab, Dept Biochem, S-10691 Stockholm, Sweden
[2] Slovak Acad Sci, Canc Res Inst, SK-83391 Bratislava, Slovakia
[3] Charles Univ, Fac Nat Sci, Dept Physiol, Prague 12000 3, Czech Republic
关键词
DNase I protection; promoter regulation; repressor; transcription;
D O I
10.1042/0264-6021:3520519
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Regulatory factors bound to the human adenine nucleotide translocator-2 (ANT2) promoter have been mapped in HeLa cells by in vivo DNase I protection and ligation-mediated PCR amplification. Protein binding was detected at only three sites within the extended promoter region (to nt - 703). One, starting at nt -61 and covering the TATA box and transcription start, most probably represents occupation by the transcription-initiation machinery. A repeated Spl element determined by in vitro studies to be the major activation element for the promoter was also protected in vivo on nucleotides responsible for strong binding to the zinc fingers. Occupation of two additional upstream Sp1 elements was not observed. The third site occupied in vivo was identified previously by in vitro studies as a unique silencer element. Treatment of cells with trichostatin A to induce hyperacetylation released the silencer-binding protein after 1 h, but had no effect on the Sp1-activating elements. Prolonged treatment (24 h) displaced Sp1 from the activating elements. These findings confirm and extend in vitro studies indicating that regulation of the ANT2 promoter is most probably exerted through a single pair of proximal Sp1-activating elements and an upstream silencer, and that chromatin organization plays a role in the interaction between the two.
引用
收藏
页码:519 / 523
页数:5
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