Detecting protein-protein interactions in vesicular stomatitis virus using a cytoplasmic yeast two hybrid system.
Detecting protein-protein interactions in vesicular stomatitis virus using a cytoplasmic yeast two hybrid system.
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DOI：
10.1016/j.jviromet.2011.02.006
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发表时间：
2011-05
影响因子：
3.1
通讯作者：
Grdzelishvili, Valery Z.
中科院分区：
文献类型：
--
作者：
Moerdyk-Schauwecker, Megan;DeStephanis, Darla;Hastie, Eric;Grdzelishvili, Valery Z.
关键词：
蛋白质相互作用在病毒编码的许多功能和病毒与宿主相互作用中起着重要作用。虽然“经典”酵母双杂交系统（Y2H）是检测这种相互作用的最常见技术之一，但它有许多限制，包括需要将感兴趣的蛋白质重新定位到细胞核。修饰的Y2H，如Sos募集系统（SRS），检测发生在细胞质而不是细胞核中的相互作用，允许在更自然的环境中测试来自细胞质中复制的病毒的蛋白质。在这项研究中，SRS被用来检测涉及水泡性口炎病毒（VSV），一个原型的非节段负链RNA（NNS）病毒的蛋白质的相互作用。所有五种全长VSV蛋白以及几种截短的蛋白相互筛选。使用SRS，大多数相互作用先前证明涉及VSV磷蛋白，核衣壳（N）和大聚合酶蛋白被独立确认，而遇到的困难，使用膜相关的基质和糖蛋白。还针对VSV N蛋白筛选了人cDNA文库，并鉴定了一种与N相互作用的细胞蛋白SFRS 18。所提出的系统可以很容易地重新设计，在其他不太听话的NNS病毒的研究。
Protein-protein interactions play an important role in many virus-encoded functions and in virus-host interactions. While a “classical” yeast two-hybrid system (Y2H) is one of the most common techniques to detect such interactions, it has a number of limitations, including a requirement for the proteins of interest to be relocated to the nucleus. Modified Y2H, such as the Sos recruitment system (SRS), which detect interactions occurring in the cytoplasm rather than the nucleus, allow proteins from viruses replicating in the cytoplasm to be tested in a more natural context. In this study, a SRS was used to detect interactions involving proteins from vesicular stomatitis virus (VSV), a prototypic non-segmented negative strand RNA (NNS) virus. All five full-length VSV proteins, as well as several truncated proteins, were screened against each other. Using the SRS, most interactions demonstrated previously involving VSV phosphoprotein, nucleocapsid (N) and large polymerase proteins were confirmed independently, while difficulties were encountered using the membrane associated matrix and glycoproteins. A human cDNA library was also screened against VSV N protein and one cellular protein, SFRS18, was identified which interacted with N in this context. The system presented can be redesigned easily for studies in other less tractable NNS viruses.
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影响因子：
--
作者：
Broach, J R;Deschenes, R J
通讯作者：
Deschenes, R J
影响因子：
4.8
作者：
Chou, Han-Yi E.;Chou, Han Ting;Lee, Sheng-Chung
通讯作者：
Lee, Sheng-Chung
影响因子：
6.7
作者：
Fossum E;Friedel CC;Rajagopala SV;Titz B;Baiker A;Schmidt T;Kraus T;Stellberger T;Rutenberg C;Suthram S;Bandyopadhyay S;Rose D;von Brunn A;Uhlmann M;Zeretzke C;Dong YA;Boulet H;Koegl M;Bailer SM;Koszinowski U;Ideker T;Uetz P;Zimmer R;Haas J
通讯作者：
Haas J
影响因子：
3.7
作者：
Barak, O;Aronheim, A;Shaul, Y
通讯作者：
Shaul, Y
DOI：
10.1073/pnas.83.23.8873
发表时间：
1986-12-01
影响因子：
11.1
作者：
GILL, DS;CHATTOPADHYAY, D;BANERJEE, AK
通讯作者：
BANERJEE, AK