Virion-Packaged Pyruvate Kinase Muscle Type 2 Affects Reverse Transcription Efficiency of Human Immunodeficiency Virus Type 1 by Blocking Virion Recruitment of tRNALys3.-猫眼课题宝

病毒粒子包装的丙酮酸激酶肌肉 2 型通过阻止病毒粒子招募 tRNALys3 来影响人类免疫缺陷病毒 1 型的逆转录效率。

DOI：

发表时间：

2018

期刊：

Biological and Pharmaceutical Bulletin

影响因子：

通讯作者：

中科院分区：

医学4区

文献类型：

作者：

Kumkum Rahman Mouree;N. Kishimoto;Nozomi Iga;Chie Kirihara;Kengo Yamamoto;N. Takamune;S. Misumi

人类免疫缺陷病毒 1 型 (HIV-1) 在其形态发生过程中将多种细胞因子招募到病毒颗粒中，这显然在调节其感染性方面发挥了作用。在我们的研究中，蛋白质组学技术证明，一种关键的糖酵解蛋白，丙酮酸激酶肌肉 2 型 (PKM2)，已整合到病毒颗粒中。在这里，我们表明，病毒颗粒包装的 PKM2 通过影响细胞 tRNALys3 掺入病毒颗粒的水平，显着降低病毒感染性。产生 HIV-1 的细胞中 PKM2 表达的增强导致 PKM2 掺入子代病毒粒子的水平更高，而不影响病毒成熟过程。与对照病毒相比，高水平PKM2包装病毒的逆转录产物和细胞tRNALys3包装水平均降低，表明病毒颗粒内tRNALys3的缺乏抑制了靶细胞中的逆转录效率。有趣的是，PKM2 表达的增强还抑制了其他非引发细胞 tRNA（例如 tRNALys1,2 和 tRNAAsn）的病毒体招募，已知这些 tRNA 被选择性包装到病毒体中，而不影响生产细胞中这些 tRNA 的细胞质库的稳定水平，这表明 PKM2 特别阻碍 tRNA 选择性掺入病毒体。综上所述，我们的研究结果表明 PKM2 是一个重要的宿主因子，它通过靶向靶细胞中 tRNALys3 介导的逆转录起始，对 HIV-1 感染性产生负面影响。

Human immunodeficiency virus type 1 (HIV-1) recruits diverse cellular factors into viral particles during its morphogenesis, which apparently play roles in modulating its infectivity. In our study, proteomic techniques demonstrated that a key glycolytic protein, pyruvate kinase muscle type 2 (PKM2), is incorporated into viral particles. Here, we show that virion-packaged PKM2 significantly reduces viral infectivity by affecting the incorporation level of a cellular tRNALys3 into virions. Enhanced expression of PKM2 in HIV-1-producing cells led to a higher incorporation level of PKM2 into progeny virions without affecting the viral maturation process. Compared with the control virus, the high-level-PKM2-packaging virus showed decreased levels of both reverse transcription products and cellular tRNALys3 packaging, suggesting that the shortage of intravirion tRNALys3 suppresses reverse transcription efficiency in target cells. Interestingly, the enhanced expression of PKM2 also suppressed the virion recruitment of other nonpriming cellular tRNAs such as tRNALys1,2 and tRNAAsn, which are known to be selectively packaged into virions, without affecting the steady level of the cytoplasmic pool of those tRNAs in producer cells, suggesting that PKM2 specifically impedes the selective incorporation of tRNAs into virions. Taken together, our findings indicate that PKM2 is a vital host factor that negatively affects HIV-1 infectivity by targeting the tRNALys3-mediated initiation of reverse transcription in target cells.

Human immunodeficiency virus type 1 Nef and p56lck protein-tyrosine kinase interact with a common element in CD4 cytoplasmic tail.