Protection against fatal Sindbis virus encephalitis by Beclin, a novel Bcl-2-interacting protein
Protection against fatal Sindbis virus encephalitis by Beclin, a novel Bcl-2-interacting protein
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DOI：
10.1128/jvi.72.11.8586-8596.1998
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发表时间：
1998-11-01
影响因子：
5.4
通讯作者：
Levine, B
中科院分区：
文献类型：
--
作者：
Liang, XH;Kleeman, LK;Levine, B
原型细胞抗凋亡基因bcl-2可减少Sindbis病毒在小鼠脑中的复制和Sindbis病毒诱导的细胞凋亡，从而保护小鼠免受致死性脑炎的侵袭。为了研究Bcl-2抵抗中枢神经系统Sindbis病毒感染的潜在机制，我们在成年小鼠脑文库中进行酵母双杂交筛选，以鉴定Bcl-2相互作用的基因产物。为了研究Beclin在Sindbis病毒发病机制中的作用，我们构建了重组Sindbis病毒嵌合体，表达全长人Beclin （SIN/ Beclin）、缺乏Bcl-2结合结构域的Beclin （SIN/ Beclin Delta Bcl-2BD）或在5'端附近含有过早终止密码子的Beclin （SIN/beclinstop）。与SIN/beclin δ Bcl-2BD或SIN/beclinstop相比，SIN/beclin δ Bcl-2BD感染小鼠的存活率（71%）显著高于SIN/beclin δ Bcl-2BD感染小鼠的存活率（9%）和SIN/beclinstop感染小鼠的存活率（7%）（P < 0.001）， SIN/beclin δ Bcl-2BD感染小鼠的大脑中Sindbis病毒rna阳性细胞较少，凋亡细胞较少，病毒滴度也较SIN/beclinstop感染小鼠的大脑低。这些发现表明Beclin是一种新的bcl -2相互作用的细胞蛋白，可能在抗病毒宿主防御中发挥作用。
bcl-2, the prototypic cellular antiapoptotic gene, decreases Sindbis virus replication and Sindbis virus-induced apoptosis in mouse brains, resulting in protection against lethal encephalitis. To investigate potential mechanisms by which Bcl-2 protects against central nervous system Sindbis virus infection, we performed a yeast two-hybrid screen to identify Bcl-2-interacting gene products in an adult mouse brain library. We identified a novel 60-kDa coiled-coil protein, Beelin, which we confirmed interacts with Bcl-2 in mammalian cells, using fluorescence resonance energy transfer microscopy, To examine the role of Beclin in Sindbis virus pathogenesis, we constructed recombinant Sindbis virus chimeras that express full-length human Beclin (SIN/beclin), Beclin lacking the putative Bcl-2-binding domain (SIN/beclin Delta Bcl-2BD), or Beclin containing a premature stop codon near the 5' terminus (SIN/beclinstop). The survival of mice infected with SIN/beclin was significantly higher (71%) than the survival of mice infected with SIN/beclin Delta Bcl-2BD (9%) or SIN/beclinstop (7%) (P < 0.001), The brains of mice infected with SIN/beclin had fewer Sindbis virus RNA-positive cells, fewer apoptotic cells, and lower viral titers than the brains of mice infected with SIN/beclin Delta Bcl-2BD or SIN/beclinstop, These findings demonstrate that Beclin is a novel Bcl-2-interacting cellular protein that may play a role in antiviral host defense.