IDENTIFICATION AND CHARACTERIZATION OF A HELA NUCLEAR-PROTEIN THAT SPECIFICALLY BINDS TO THE TRANS-ACTIVATION-RESPONSE (TAR) ELEMENT OF HUMAN-IMMUNODEFICIENCY-VIRUS

被引:216
作者
MARCINIAK, RA
GARCIABLANCO, MA
SHARP, PA
机构
[1] MIT,CTR CANC RES,CAMBRIDGE,MA 02139
[2] MIT,DEPT BIOL,CAMBRIDGE,MA 02139
[3] HARVARD UNIV,MIT,DIV HLTH SCI & TECHNOL,CAMBRIDGE,MA 02139
关键词
RNA-binding proteins; UV cross-linking;
D O I
10.1073/pnas.87.9.3624
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
Human immunodeficiency virus type 1 RNAs contain a sequence, trans-activation-response (TAR) element, which is required for tat protein-mediated trans-activation of viral gene expression. We have identified a nuclear protein from extracts of HeLa cells that binds to the TAR element RNA in a sequence-specific manner. The binding of this 68-kDa polypeptide was detected by UV cross-linking proteins to TAR element RNA transcribed in vitro. Competition experiments were performed by using a partially purified preparation of the protein to quantify the relative binding affinities of TAR element RNA mutants. The binding affinity of the TAR mutants paralleled the reported ability of those mutants to support tat trans-activation in vivo. We propose that this cellular protein moderates TAR activity in vivo.
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页码:3624 / 3628
页数:5
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