title: Two second-site mutations compensate the engineered mutation of R7A in vesicular stomatitis virus nucleocapsid protein.
abstract: The functional template for the transcription and replication of vesicular stomatitis virus (VSV) is genomic RNA encapsidated by nucleocapsid (N) protein. Previous studies showed that the amino acid R7 in the N-terminal arm of N is involved in N-N interaction in the N-RNA complex. In our study, the recombinant virus with mutation of R7A (rVSV(R7A)) in N was recovered, and the replication level of passage 1 (P1) of rVSV(R7A) was 1000 times lower than that of wild-type rVSV at 37°C. After eight passages, the replication level of P8 of rVSV(R7A) with two second-site mutations in the genome (T242 P in N protein and U7-U8 in G-L gene junction) was significantly higher than that of P1. Furthermore, we demonstrate that the mutation of either T242P or U7-U8 can compensate the effect caused by the mutation of R7A on the replication of rVSV(R7A). Therefore, we conclude that two second-site mutations both can compensated the engineered mutation of R7A in VSV N protein.
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, China; Hubei University of Chinese Medicine, Basic medical college, Wuhan, China.
authors.forename: Longyun
authors.initials: L
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhong
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, China.
authors.forename: Zhulong
authors.initials: Z
authors.lastname: Hu
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, China.
authors.forename: Yali
authors.initials: Y
authors.lastname: Qin
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, China. Electronic address: chenmz@whu.edu.cn.
authors.forename: Mingzhou
authors.initials: M
authors.lastname: Chen
mesh_terms: D005816:Genetic Complementation Test; D005818:Genetic Engineering; D016679:Genome, Viral; D008958:Models, Molecular; D009154:Mutation; D019590:Nucleocapsid Proteins / Q000235:genetics*; D011485:Protein Binding; D011487:Protein Conformation; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics* / Q000502:physiology; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics; D014779:Virus Replication
chemical_list: D019590:Nucleocapsid Proteins; D012367:RNA, Viral; D014764:Viral Proteins