Characterization and modification of the carboxy-terminal sequences of bluetongue virus type 10 NS1 protein in relation to tubule formation and location of an antigenic epitope in the vicinity of the carboxy terminus of the protein.
Characterization and modification of the carboxy-terminal sequences of bluetongue virus type 10 NS1 protein in relation to tubule formation and location of an antigenic epitope in the vicinity of the carboxy terminus of the protein.
复制标题
蓝舌病毒 10 型 NS1 蛋白羧基末端序列的表征和修饰与小管形成和蛋白羧基末端附近抗原表位位置的关系。
DOI：
10.1128/jvi.69.5.2831-2841.1995
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发表时间：
1995
影响因子：
5.4
通讯作者：
Roy,P
中科院分区：
文献类型：
--
作者：
Monastyrskaya,K;Gould,EA;Roy,P
蓝舌病毒在感染过程中会产生大量的小管。这些小管是由病毒双链核糖核酸片段M6编码的552个氨基酸的-kDa NS1蛋白形成的。蓝舌病毒血清10型NS1的一系列缺失和延伸突变体已在昆虫细胞中表达，以鉴定该蛋白中对小管形成至关重要的羧基末端成分。分别制备了缺失5个和10个羧基末端残基的突变体AcCT5和AcCT10。通过分析它们形成小管的能力，发现AcCT5具有这一功能，而AcCT10不具有这一功能，这表明后5个氨基酸在NS1小管形成过程中没有很强的参与。含有外源抗原序列的延伸突变体在NS1的C末端增加了多达16个氨基酸，形成了小管，但19个氨基酸的延伸抑制了小管的形成。对一组单抗的分析已经确定，NS1抗原点位于蛋白质的羧基末端附近。它似乎暴露在小管的表面。讨论了利用重组NS1传递外源表位来开发新疫苗的机会。
Bluetongue virus produces large numbers of tubules during infection. The tubules are formed from a 552-amino-acid, 64-kDa NS1 protein encoded by the viral double-stranded RNA segment M6. A series of deletion and extension mutants of bluetongue virus serotype 10 NS1 has been generated and expressed in insect cells in order to identify the carboxy-terminal components of the protein which are important for tubule formation. The mutants AcCT5 and AcCT10, lacking 5 and 10 of the carboxy-terminal residues, respectively, were prepared. By analyzing their abilities to form tubules, it was shown that AcCT5 was capable of this function whereas AcCT10 was not, indicating that the last five amino acids are not strongly involved in NS1 tubule formation. Extension mutants including foreign antigenic sequences involving up to 16 amino acids added to the C terminus of NS1 were shown to form tubules, although an extension of 19 amino acids inhibited tubule formation. Analysis of a panel of monoclonal antibodies has established that an NS1 antigenic site is located near the carboxy terminus of the protein. It appears to be exposed on the surface of tubules. The opportunities to develop new vaccines using recombinant NS1 to deliver foreign epitopes are discussed.