Interaction of Rotaviruses with Hsc70 during Cell Entry Is Mediated by VP5
Interaction of Rotaviruses with Hsc70 during Cell Entry Is Mediated by VP5
复制标题
DOI：
10.1128/jvi.77.13.7254-7260.2003
复制
发表时间：
2003-07
影响因子：
5.4
通讯作者：
S. Zárate;M. A. Cuadras;R. Espinosa;P. Romero;Karla O. Juárez;Minerva Camacho-Nuez;C. Arias;S. L
中科院分区：
文献类型：
--
作者：
S. Zárate;M. A. Cuadras;R. Espinosa;P. Romero;Karla O. Juárez;Minerva Camacho-Nuez;C. Arias;S. L
轮状病毒感染似乎是一个多步骤的过程，在此过程中，病毒需要与几种细胞表面分子相互作用才能进入细胞。病毒刺突蛋白VP4被胰蛋白酶切割成两个亚基VP5和VP8，参与了其中的一些相互作用。我们之前已经证明，神经氨酸酶敏感轮状病毒株RRV最初通过VP4的VP8亚基附着于含唾液酸的细胞分子，随后通过VP5与整合素α2β1相互作用。在这些初始接触之后，病毒与位于细胞表面的至少两种另外的蛋白质相互作用，整合素αvβ3和热休克同源蛋白Hsc 70。在这项工作中，我们已经表明，轮状病毒RRV和它的神经氨酸酶耐药变异nar3与Hsc70通过VP5结构域之间的氨基酸642和658的蛋白质。该结论基于以下观察结果：包含VP5的300个羧基末端氨基酸的重组蛋白特异性结合Hsc 70，并且包含氨基酸642至658的合成肽与RRV和nar 3病毒竞争结合热休克蛋白。VP5肽还与重组VP5蛋白质的Hsc70的结合竞争，并且Hsc70的抗体减少重组蛋白质与MA104细胞表面的结合。合成肽阻断轮状病毒RRV和nar 3的感染性但不阻断它们与细胞的结合的事实表明，VP5与Hsc 70的相互作用最可能发生在病毒进入过程中的后附着步骤。
ABSTRACT Rotavirus infection seems to be a multistep process in which the viruses are required to interact with several cell surface molecules to enter the cell. The virus spike protein VP4, which is cleaved by trypsin into two subunits, VP5 and VP8, is involved in some of these interactions. We have previously shown that the neuraminidase-sensitive rotavirus strain RRV initially attaches to a sialic acid-containing cell molecule through the VP8 subunit of VP4 and subsequently interacts with integrin α2β1 through VP5. After these initial contacts, the virus interacts with at least two additional proteins located at the cell surface, the integrin αvβ3 and the heat shock cognate protein Hsc70. In this work, we have shown that rotavirus RRV and its neuraminidase-resistant variant nar3 interact with Hsc70 through a VP5 domain located between amino acids 642 and 658 of the protein. This conclusion is based on the observation that a recombinant protein comprising the 300 carboxy-terminal amino acids of VP5 binds specifically to Hsc70 and a synthetic peptide containing amino acids 642 to 658 competes with the binding of the RRV and nar3 viruses to the heat shock protein. The VP5 peptide also competed with the binding to Hsc70 of the recombinant VP5 protein, and an antibody to Hsc70 reduced the binding of the recombinant protein to the surface of MA104 cells. The fact that the synthetic peptide blocks the infectivity of rotaviruses RRV and nar3 but not their binding to cells indicates that the interaction of VP5 with Hsc70 most probably occurs at a postattachment step during the virus entry process.