【摘要】：異嗜性鼠白血病病毒相關(guān)病毒(XMRV)是新發(fā)現(xiàn)的一種具有完全復(fù)制能力的γ逆轉(zhuǎn)錄病毒。早期研究報道XMRV可能與人類疾病前列腺癌、慢性疲勞綜合癥(CFS)有關(guān),但隨后的研究中未能在前列腺癌和慢性疲勞綜合癥的組織樣本中檢測出XMRV,甚至在早期檢出XMRV的樣本中也未能檢測出XMRV,認(rèn)為早期的研究結(jié)果是實驗過程中的污染導(dǎo)致的,排除了XMRV與這兩種人類疾病的直接關(guān)系。最新研究證實XMRV是在產(chǎn)生前列腺癌細(xì)胞系時由兩個內(nèi)源性鼠白血病病毒重組產(chǎn)生的。雖然最近的研究報道XMRV感染與人類疾病之間并沒有直接關(guān)系,但XMRV仍然是一種未定義的潛在病原體,因為在體外它仍具有感染人類細(xì)胞的能力。并且XMRV具有γ逆轉(zhuǎn)錄病毒的性質(zhì),可以作為模式病毒研究宿主細(xì)胞因子與逆轉(zhuǎn)錄病毒之間的相互作用。研究發(fā)現(xiàn)在宿主細(xì)胞中存在一些限制因子APOBEC3和Tetherin蛋白等可以抑制XMRV的復(fù)制。 宿主因子MOV10蛋白廣泛存在于人類、鼠類等多種物種中,在胚胎干細(xì)胞等多種細(xì)胞中廣泛表達(dá)。MOV10蛋白是RNA解旋酶超家族-1中的一個成員,它也是一種RNA誘導(dǎo)的沉默復(fù)合體(RISC)的組成部分。近期研究表明MOV10可以抑制包括HIV-1在內(nèi)的逆轉(zhuǎn)錄病毒的復(fù)制,具有廣譜抗逆轉(zhuǎn)錄病毒的性質(zhì),在RNA干擾途徑中起重要作用。MOV10蛋白本身具有RNA解旋酶的7個保守結(jié)構(gòu)域,在MOV10抑制逆轉(zhuǎn)錄病毒中發(fā)揮重要作用。 本研究的內(nèi)容是研究MOV10抑制XMRV復(fù)制的作用及其抑制作用機(jī)制,探索MOV10抑制活性的關(guān)鍵區(qū)域,具有一定的創(chuàng)新性。 本研究采用XMRV前病毒DNA質(zhì)粒與MOV10質(zhì)粒/MOV10突變體共轉(zhuǎn)染哺乳動物細(xì)胞,產(chǎn)出的病毒再感染細(xì)胞,通過檢測病毒感染性的方法來驗證MOV10對XMRV的抑制作用及其機(jī)制以及MOV10對XMRV發(fā)揮抑制作用的關(guān)鍵區(qū)域。 通過本研究得出以下結(jié)論：MOV10具有抑制XMRV復(fù)制的活性,過量表達(dá)MOV10時,XMRV的感染性顯著降低,而降低內(nèi)源性MOV10的表達(dá)時,XMRV的感染性明顯增強(qiáng)。MOV10通過包裝進(jìn)XMRV病毒顆粒中,從而在病毒再感染細(xì)胞時發(fā)揮抑制作用,MOV10發(fā)揮抑制作用的主要階段是XMRV生活周期的逆轉(zhuǎn)錄階段。MOV10發(fā)揮抑制作用的關(guān)鍵區(qū)域主要是MOV10本身的RNA解旋酶的7個保守結(jié)構(gòu)域,其中結(jié)構(gòu)域Ⅰ、Ⅱ、Ⅲ和Ⅳ的作用非常重要,結(jié)構(gòu)域V和Ⅵ的作用其次,結(jié)構(gòu)域Ia的作用最小。 本研究的成果證明了MOV10可以抑制XMRV的復(fù)制,初步闡明了MOV10抑制XMRV復(fù)制的機(jī)制,明確了MOV10發(fā)揮抑制作用的關(guān)鍵結(jié)構(gòu)域。進(jìn)一步證實了MOV10具有廣譜的抗逆轉(zhuǎn)錄病毒的活性,在宿主抵抗逆轉(zhuǎn)錄病毒感染中發(fā)揮了積極重要的作用。
[Abstract]:Heterophil leukemia virus associated virus (XMRV) is a newly discovered 緯-retrovirus with complete replication ability. Early studies have reported that XMRV may be associated with prostate cancer and chronic fatigue syndrome (CFS) in humans, but subsequent studies failed to detect XMRV, in tissue samples of prostate cancer and chronic fatigue syndrome. Even in the early detection of XMRV samples, XMRV, could not detect that the early results of the study were caused by pollution in the experimental process, excluding the direct relationship between XMRV and these two human diseases. New research confirms that XMRV is produced by recombinant two endogenous murine leukemia viruses when prostate cancer cell lines are produced. Although recent studies have reported that there is no direct relationship between XMRV infection and human disease, XMRV is still an undefined potential pathogen because it still has the ability to infect human cells in vitro. Moreover, XMRV has the property of 緯 retrovirus and can be used as a model virus to study the interaction between host cytokines and retroviruses. It was found that some limiting factors, such as APOBEC3 and Tetherin proteins, could inhibit the replication of XMRV in host cells. Host factor MOV10 protein is widely expressed in human, mouse and other species, and is widely expressed in embryonic stem cells and other cells. MOV10 protein is a member of RNA helicase superfamily 1. It is also part of a RNA-induced silencing complex (RISC). Recent studies have shown that MOV10 can inhibit the replication of retroviruses, including HIV-1, and has the properties of broad-spectrum antiretrovirals and plays an important role in the RNA interference pathway. The MOV10 protein itself has seven conserved domains of RNA helicase. It plays an important role in the inhibition of retrovirus by MOV10. The purpose of this study is to study the inhibitory effect of MOV10 on XMRV replication and its mechanism, and to explore the key regions of MOV10 inhibitory activity. In this study, mammalian cells were co-transfected with XMRV precursor virus DNA plasmid and MOV10 plasmid / MOV10 mutants to produce virus reinfection cells. The inhibition of XMRV by MOV10 and its mechanism and the key regions of XMRV inhibition by MOV10 were verified by the method of virus infection. The results showed that MOV10 had the activity of inhibiting XMRV replication. When MOV10 was over-expressed, the infectivity of XMRV was significantly decreased, while that of XMRV was significantly enhanced when the expression of endogenous MOV10 was decreased. MOV10 was packaged into XMRV virus particles. The inhibitory effect of MOV10 is the reverse transcription stage of XMRV life cycle. The key region of inhibition of MOV10 is the seven conserved domains of RNA helicase of MOV10 itself. Domain 鈪，

本文編號：2329648