Human immunodeficiency virus type-1 Tat is an integral component of the activated transcription-elongation complex

被引:88
|
作者
Keen, NJ [1 ]
Gait, MJ [1 ]
Karn, J [1 ]
机构
[1] MRC,MOLEC BIOL LAB,CAMBRIDGE CB2 2QH,ENGLAND
关键词
RNA polymerase II; transactivation;
D O I
10.1073/pnas.93.6.2505
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The human immunodeficiency virus type 1 transactivator protein, Tat, stimulates transcriptional elongation from the viral long terminal repeat, To test whether Tat associates directly with activated transcription complexes, we have used the lac repressor protein (LacR) to ''trap'' elongating RNA polymerases. The arrested transcription complexes were purified by binding biotinylated templates to streptavidin-coated magnetic beads. Transcription complexes were released from the magnetic beads following cleavage of the templates with restriction enzymes and were immunoblotted with antibodies to Tat, LacR and RNA polymerase II, The Tat protein copurified with RNA polymerase bound to wild-type templates but did not copurify with transcription complexes prepared by using templates carrying mutations in the transactivation response element (TAR) RNA, We conclude that Tat and cellular cofactors become attached to the transcription complex during its transit through TAR.
引用
收藏
页码:2505 / 2510
页数:6
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