The interaction of LEDGF/p75 with integrase is lentivirus-specific and promotes DNA binding
The interaction of LEDGF/p75 with integrase is lentivirus-specific and promotes DNA binding
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DOI：
10.1074/jbc.m411681200
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发表时间：
2005-05-06
影响因子：
4.8
通讯作者：
Debyser, Z
中科院分区：
文献类型：
--
作者：
Busschots, K;Vercammen, J;Debyser, Z
我们先前已经表明，转录共激活因子透镜上皮衍生生长因子（LEDGF）的p75同种型与人免疫缺陷病毒（HIV）-1整合酶（IN）紧密相互作用，并且对于该蛋白质在人细胞中的核靶向是必需的（Cherepanov，P.，Maertens，G.，Proost，P.，Devreese，B.，货车Beeumen，J.，Engelborghs，Y.，德克莱克，E.，Debyser，Z.（2003）J.Biol.Chem.278，372 - 381; Maertens，G.，切列帕诺夫，P。，Pluymers，W.，Busschots，K.，德克莱克，E.，Debyser，Z.，和Engelborghs，Y.（2003）J.Biol.Chem.278，33528 - 33539）。在此，在下拉结合试验中检查重组LEDGF/p75和HIV-1 IN之间的相互作用。LEDGF/p75显示增加HIV-1 IN的溶解度。接着，使用荧光相关光谱法测量LEDGF/p75或HIV-1 IN和LEDGF/p75的复合物与特异性双链DNA寡核苷酸的相互作用。尽管LEDGF/p75对DNA仅显示中等亲和力，但它强烈促进HIV-1 IN与DNA的结合。这种效应对于LEDGF的p75同种型是特异性的，而对于p52则没有观察到。在下拉试验中，LEDGF/p75与HIV-1、HIV-2和猫免疫缺陷病毒IN相互作用，但不与人T细胞嗜淋巴细胞病毒2型、莫洛尼鼠白血病病毒或劳斯肉瘤病毒的IN相互作用。这些结果强烈表明LEDGF/p75与IN的相互作用对慢病毒科是特异性的。LEDGF/ p75刺激HIV-1和HIV-2 IN与非特异性DNA的结合，但不刺激Moloney鼠白血病病毒或Rous肉瘤病毒IN。这些结果为我们的假设提供了支持证据，即LEDGF/ p75在慢病毒IN与染色体DNA的连接中起作用。
We have previously shown that the p75 isoform of the transcriptional co-activator lens epithelium-derived growth factor ( LEDGF) interacts tightly with human immunodeficiency virus (HIV)-1 integrase ( IN) and is essential for nuclear targeting of this protein in human cells (Cherepanov, P., Maertens, G., Proost, P., Devreese, B., Van Beeumen, J., Engelborghs, Y., De Clercq, E., and Debyser, Z. ( 2003) J. Biol. Chem. 278, 372 - 381; Maertens, G., Cherepanov, P., Pluymers, W., Busschots, K., De Clercq, E., Debyser, Z., and Engelborghs, Y. ( 2003) J. Biol. Chem. 278, 33528 - 33539). Here the interaction between recombinant LEDGF/p75 and HIV-1 IN was examined in a pull-down binding test. LEDGF/p75 was shown to increase the solubility of HIV-1 IN. Next, fluorescent correlation spectroscopy was used to measure the interaction of LEDGF/p75 or the complex of HIV-1 IN and LEDGF/p75 with a specific double-stranded DNA oligonucleotide. Whereas LEDGF/p75 displayed only a moderate affinity for DNA, it strongly promoted the binding of HIV-1 IN to DNA. This effect was specific for the p75 isoform of LEDGF and was not seen with p52. In the pull-down assay LEDGF/p75 interacted with HIV-1, HIV-2, and feline immunodeficiency virus IN, but not with the IN of human T-cell lymphotropic virus type 2, Moloney murine leukemia virus, or Rous sarcoma virus. These results strongly suggest that the interaction of LEDGF/p75 with IN is specific to lentiviridae. LEDGF/ p75 stimulated the binding of HIV-1 and HIV-2 IN, but not Moloney murine leukemia virus or Rous sarcoma virus IN, to an aspecific DNA. These results provide supporting evidence for our hypothesis that LEDGF/ p75 plays a role in the tethering of lentiviral IN to the chromosomal DNA.