Alteration of Sendai virus morphogenesis and nucleocapsid incorporation due to mutation of cysteine residues of the matrix protein

Takemasa Sakaguchi, Tsuneo Uchiyama, Cheng Huang, Noriko Fukuhara, Katsuhiro Kiyotani, Yoshiyuki Nagai, Tetsuya Yoshida

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

The matrix (M) protein of Sendai virus (SeV) has five cysteine residues, at positions 83, 106, 158, 251, and 295. To determine the roles of the cysteine residues in viral assembly, we generated mutant M cDNA possessing a substitution to serine at one of the cysteine residues or at all of the cysteine residues. Some mutant M proteins were unstable when expressed in cultured cells, suggesting that cysteine residues affect protein stability, probably by disrupting the proper conformation. In an attempt to generate virus from cDNA, SeV M-C83S, SeV M-C106S, and SeV M-C295S were successfully recovered from cDNA, while recombinant SeVs possessing other mutations were not. SeV M-C83S and SeV M-C106S had smaller virus particles than did the wild-type SeV, whereas SeV M-C295S had larger and heterogeneously sized particles. Furthermore, SeV M-C106S had a significant amount of empty particles lacking nucleocapsids. These results indicate that a single-point mutation at a cysteine residue of the M protein affects virus morphology and nucleocapsid incorporation, showing direct involvement of the M protein in SeV assembly. Cysteine-dependent conformation of the M protein was not due to disulfide bond formation, since the cysteines were shown to be free throughout the viral life cycle.

Original languageEnglish (US)
Pages (from-to)1682-1690
Number of pages9
JournalJournal of Virology
Volume76
Issue number4
DOIs
StatePublished - Feb 5 2002
Externally publishedYes

Fingerprint

Sendai virus
Nucleocapsid
nucleocapsid
Morphogenesis
morphogenesis
Cysteine
cysteine
mutation
Mutation
Proteins
proteins
Virus Assembly
Complementary DNA
Viruses
Protein Conformation
Protein Stability
Mutant Proteins
Life Cycle Stages
viruses
mutants

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Alteration of Sendai virus morphogenesis and nucleocapsid incorporation due to mutation of cysteine residues of the matrix protein. / Sakaguchi, Takemasa; Uchiyama, Tsuneo; Huang, Cheng; Fukuhara, Noriko; Kiyotani, Katsuhiro; Nagai, Yoshiyuki; Yoshida, Tetsuya.

In: Journal of Virology, Vol. 76, No. 4, 05.02.2002, p. 1682-1690.

Research output: Contribution to journalArticle

Sakaguchi, Takemasa ; Uchiyama, Tsuneo ; Huang, Cheng ; Fukuhara, Noriko ; Kiyotani, Katsuhiro ; Nagai, Yoshiyuki ; Yoshida, Tetsuya. / Alteration of Sendai virus morphogenesis and nucleocapsid incorporation due to mutation of cysteine residues of the matrix protein. In: Journal of Virology. 2002 ; Vol. 76, No. 4. pp. 1682-1690.
@article{3039f720a55d4dea98fd614861e49254,
title = "Alteration of Sendai virus morphogenesis and nucleocapsid incorporation due to mutation of cysteine residues of the matrix protein",
abstract = "The matrix (M) protein of Sendai virus (SeV) has five cysteine residues, at positions 83, 106, 158, 251, and 295. To determine the roles of the cysteine residues in viral assembly, we generated mutant M cDNA possessing a substitution to serine at one of the cysteine residues or at all of the cysteine residues. Some mutant M proteins were unstable when expressed in cultured cells, suggesting that cysteine residues affect protein stability, probably by disrupting the proper conformation. In an attempt to generate virus from cDNA, SeV M-C83S, SeV M-C106S, and SeV M-C295S were successfully recovered from cDNA, while recombinant SeVs possessing other mutations were not. SeV M-C83S and SeV M-C106S had smaller virus particles than did the wild-type SeV, whereas SeV M-C295S had larger and heterogeneously sized particles. Furthermore, SeV M-C106S had a significant amount of empty particles lacking nucleocapsids. These results indicate that a single-point mutation at a cysteine residue of the M protein affects virus morphology and nucleocapsid incorporation, showing direct involvement of the M protein in SeV assembly. Cysteine-dependent conformation of the M protein was not due to disulfide bond formation, since the cysteines were shown to be free throughout the viral life cycle.",
author = "Takemasa Sakaguchi and Tsuneo Uchiyama and Cheng Huang and Noriko Fukuhara and Katsuhiro Kiyotani and Yoshiyuki Nagai and Tetsuya Yoshida",
year = "2002",
month = "2",
day = "5",
doi = "10.1128/JVI.76.4.1682-1690.2002",
language = "English (US)",
volume = "76",
pages = "1682--1690",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "4",

}

TY - JOUR

T1 - Alteration of Sendai virus morphogenesis and nucleocapsid incorporation due to mutation of cysteine residues of the matrix protein

AU - Sakaguchi, Takemasa

AU - Uchiyama, Tsuneo

AU - Huang, Cheng

AU - Fukuhara, Noriko

AU - Kiyotani, Katsuhiro

AU - Nagai, Yoshiyuki

AU - Yoshida, Tetsuya

PY - 2002/2/5

Y1 - 2002/2/5

N2 - The matrix (M) protein of Sendai virus (SeV) has five cysteine residues, at positions 83, 106, 158, 251, and 295. To determine the roles of the cysteine residues in viral assembly, we generated mutant M cDNA possessing a substitution to serine at one of the cysteine residues or at all of the cysteine residues. Some mutant M proteins were unstable when expressed in cultured cells, suggesting that cysteine residues affect protein stability, probably by disrupting the proper conformation. In an attempt to generate virus from cDNA, SeV M-C83S, SeV M-C106S, and SeV M-C295S were successfully recovered from cDNA, while recombinant SeVs possessing other mutations were not. SeV M-C83S and SeV M-C106S had smaller virus particles than did the wild-type SeV, whereas SeV M-C295S had larger and heterogeneously sized particles. Furthermore, SeV M-C106S had a significant amount of empty particles lacking nucleocapsids. These results indicate that a single-point mutation at a cysteine residue of the M protein affects virus morphology and nucleocapsid incorporation, showing direct involvement of the M protein in SeV assembly. Cysteine-dependent conformation of the M protein was not due to disulfide bond formation, since the cysteines were shown to be free throughout the viral life cycle.

AB - The matrix (M) protein of Sendai virus (SeV) has five cysteine residues, at positions 83, 106, 158, 251, and 295. To determine the roles of the cysteine residues in viral assembly, we generated mutant M cDNA possessing a substitution to serine at one of the cysteine residues or at all of the cysteine residues. Some mutant M proteins were unstable when expressed in cultured cells, suggesting that cysteine residues affect protein stability, probably by disrupting the proper conformation. In an attempt to generate virus from cDNA, SeV M-C83S, SeV M-C106S, and SeV M-C295S were successfully recovered from cDNA, while recombinant SeVs possessing other mutations were not. SeV M-C83S and SeV M-C106S had smaller virus particles than did the wild-type SeV, whereas SeV M-C295S had larger and heterogeneously sized particles. Furthermore, SeV M-C106S had a significant amount of empty particles lacking nucleocapsids. These results indicate that a single-point mutation at a cysteine residue of the M protein affects virus morphology and nucleocapsid incorporation, showing direct involvement of the M protein in SeV assembly. Cysteine-dependent conformation of the M protein was not due to disulfide bond formation, since the cysteines were shown to be free throughout the viral life cycle.

UR - http://www.scopus.com/inward/record.url?scp=0036146301&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036146301&partnerID=8YFLogxK

U2 - 10.1128/JVI.76.4.1682-1690.2002

DO - 10.1128/JVI.76.4.1682-1690.2002

M3 - Article

VL - 76

SP - 1682

EP - 1690

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 4

ER -