Mining the Human Complexome Database Identifies RBM14 as an XPO1-Associated Protein Involved in HIV-1 Rev Function
Mining the Human Complexome Database Identifies RBM14 as an XPO1-Associated Protein Involved in HIV-1 Rev Function
复制标题
DOI：
10.1128/jvi.03232-14
复制
发表时间：
2015-04-01
影响因子：
5.4
通讯作者：
Rice, Andrew P.
中科院分区：
文献类型：
--
作者：
Budhiraja, Sona;Liu, Hongbing;Rice, Andrew P.
通过募集宿主蛋白XPO1 (CRM1)， HIV-1 Rev蛋白介导不完全剪接病毒转录物的核输出。我们从最近描述的人类核复合体中挖掘数据，以确定与XPO1和Rev相关并参与Rev功能的宿主蛋白RBM14。使用依赖于Rev的p24报告质粒，我们发现RBM14缺失降低了Rev活性和Rev介导的细胞质中未剪接病毒转录物水平的增强。RBM14缺失也降低了病毒感染过程中p24的表达，表明RBM14限制了Rev的功能。RBM14先前已被证明定位于核副斑，核副斑是一种与保留未剪接的HIV-1转录本有关的结构，用于rev介导的核输出或降解。研究人员发现，缺失长链非编码RNA NEAT1，可使过表达的RBM14丧失增强Rev功能的能力，表明RBM14功能依赖于旁散斑完整性。我们的研究扩展了已知的Rev和XPO1的宿主细胞相互作用组，并进一步证实了paraspeckles在Rev的作用机制中的关键作用。我们的研究还验证了核络合体是一个可以从中挖掘病毒辅因子的数据库。这项研究挖掘了一个核蛋白复合物的数据库，以鉴定一种名为RBM14的细胞蛋白，该蛋白与XPO1 （CRM1）相关，XPO1是一种与HIV-1 Rev蛋白结合并介导不完全剪接病毒rna的核输出的核蛋白。功能分析表明，RBM14（一种存在于细胞核旁斑结构中的蛋白）参与了HIV-1 Rev的功能。这项研究验证了核复合物数据库作为一个参考，可以挖掘识别病毒辅因子。
By recruiting the host protein XPO1 (CRM1), the HIV-1 Rev protein mediates the nuclear export of incompletely spliced viral transcripts. We mined data from the recently described human nuclear complexome to identify a host protein, RBM14, which associates with XPO1 and Rev and is involved in Rev function. Using a Rev-dependent p24 reporter plasmid, we found that RBM14 depletion decreased Rev activity and Rev-mediated enhancement of the cytoplasmic levels of unspliced viral transcripts. RBM14 depletion also reduced p24 expression during viral infection, indicating that RBM14 is limiting for Rev function. RBM14 has previously been shown to localize to nuclear paraspeckles, a structure implicated in retaining unspliced HIV-1 transcripts for either Rev-mediated nuclear export or degradation. We found that depletion of NEAT1 RNA, a long noncoding RNA required for paraspeckle integrity, abolished the ability of overexpressed RBM14 to enhance Rev function, indicating the dependence of RBM14 function on paraspeckle integrity. Our study extends the known host cell interactome of Rev and XPO1 and further substantiates a critical role for paraspeckles in the mechanism of action of Rev. Our study also validates the nuclear complexome as a database from which viral cofactors can be mined.IMPORTANCEThis study mined a database of nuclear protein complexes to identify a cellular protein named RBM14 that is associated with XPO1 (CRM1), a nuclear protein that binds to the HIV-1 Rev protein and mediates nuclear export of incompletely spliced viral RNAs. Functional assays demonstrated that RBM14, a protein found in paraspeckle structures in the nucleus, is involved in HIV-1 Rev function. This study validates the nuclear complexome database as a reference that can be mined to identify viral cofactors.