Interaction of the adenovirus type 5 E4 Orf3 protein with promyelocytic leukemia protein isoform II is required for ND10 disruption
Interaction of the adenovirus type 5 E4 Orf3 protein with promyelocytic leukemia protein isoform II is required for ND10 disruption
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DOI：
10.1128/jvi.80.6.3042-3049.2006
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发表时间：
2006-03-01
影响因子：
5.4
通讯作者：
Leppard, KN
中科院分区：
文献类型：
--
作者：
Hoppe, A;Beech, SJ;Leppard, KN
核结构域10（ND 10）或早幼粒细胞白血病蛋白（PML）核体是球形核结构，需要PML蛋白才能形成。许多病毒在感染期间靶向这些结构。腺病毒5（Ad 5）的E4 Orf 3蛋白重排ND 10，导致PML与Orf 3共定位于核轨道或纤维中。有六种不同的PML亚型（I至VI）存在于ND 10，所有共享一个共同的N末端，但在其C末端的结构差异。在这项研究中，PML II是这六种亚型中唯一一种在体外和体内与Ad 5 E4 Orf 3直接特异性相互作用的亚型;这些结果定义了一种新的Orf 3活性。一系列的18个突变003蛋白中有3个不能与PML II相互作用，这些蛋白也不能引起ND 10重排。此外，在PML-空细胞，含有新形成的ND 10包含一个单一的PML亚型，只有ND 10形成的PML II被Orf 3重排。这些数据表明Orf 3和PML II之间的相互作用是ND 10重排发生所必需的。最后，Orf 3在体外显示出自缔合。这种活性在不能形成轨道和结合PML II的突变型Orf 3蛋白中不存在。因此，Orf 3寡聚化可能介导体内核径迹的形成，并且对于PML II结合也可能是重要的。
Nuclear domain 10 (ND10s), or promyelocytic leukemia protein (PML) nuclear bodies, are spherical nuclear structures that require PML proteins for their formation. Many viruses target these structures during infection. The E4 Orf3 protein of adenovirus 5 (Ad5) rearranges ND10s, causing PML to colocalize with Orf3 in nuclear tracks or fibers. There are six different PML isoforms (I to VI) present at ND10s, all sharing a common N terminus but with structural differences at their C termini. In this study, PML II was the only one of these six isoforms that was found to interact directly and specifically with Ad5 E4 Orf3 in vitro and in vivo; these results define a new Orf3 activity. Three of a series of 18 mutant 003 proteins were unable to interact with PML II; these were also unable to cause ND10 rearrangement. Moreover, in PML-null cells that contained neoformed ND10s comprising a single PML isoform, only ND10s formed of PML II were rearranged by Orf3. These data show that the interaction between Orf3 and PML II is necessary for ND10 rearrangement to occur. Finally, Orf3 was shown to self-associate in vitro. This activity was absent in mutant Orf3 proteins that were unable to form tracks and to bind PML II. Thus, Orf3 oligomerization may mediate the formation of nuclear tracks in vivo and may also be important for PML II binding.