title: Dissection and identification of regions required to form pseudoparticles by the interaction between the nucleocapsid (N) and membrane (M) proteins of SARS coronavirus.
abstract: When expressed in mammalian cells, the nucleocapsid (N) and membrane (M) proteins of the severe acute respiratory syndrome coronavirus (SARS-CoV) are sufficient to form pseudoparticles. To identify region(s) of the N molecule required for pseudoparticle formation, we performed biochemical analysis of the interaction of N mutants and M in HEK293 cells. Using a peptide library derived from N, we found that amino acids 101-115 constituted a novel binding site for M. We examined the ability of N mutants to interact with M and form pseudoparticles, and our observations indicated that M bound to NDelta(101-115), N1-150, N151-300, and N301-422, but not to N1-150Delta(101-115). However, pseudoparticles were formed when NDelta(101-115) or N301-422, but not N1-150 or N151-300, were expressed with M in HEK293 cells. These results indicated that the minimum portion of N required for the interaction with M and pseudoparticle formation consists of amino acids 301-422.
authors.affiliation: Department of Infectious Diseases, Research Institute, International Medical Center of Japan, Tokyo, Japan.
authors.forename: Seisuke
authors.initials: S
authors.lastname: Hatakeyama
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Matsuoka
authors.forename: Hidehiro
authors.initials: H
authors.lastname: Ueshiba
authors.forename: Nobukazu
authors.initials: N
authors.lastname: Komatsu
authors.forename: Kyogo
authors.initials: K
authors.lastname: Itoh
authors.forename: Shigeki
authors.initials: S
authors.lastname: Shichijo
authors.forename: Takao
authors.initials: T
authors.lastname: Kanai
authors.forename: Masaya
authors.initials: M
authors.lastname: Fukushi
authors.forename: Isao
authors.initials: I
authors.lastname: Ishida
authors.forename: Teruo
authors.initials: T
authors.lastname: Kirikae
authors.forename: Takehiko
authors.initials: T
authors.lastname: Sasazuki
authors.forename: Tohru
authors.initials: T
authors.lastname: Miyoshi-Akiyama
mesh_terms: D002460:Cell Line; D000086862:Coronavirus M Proteins; D000086462:Coronavirus Nucleocapsid Proteins; D019251:Nucleocapsid / Q000235:genetics / Q000378:metabolism; D019590:Nucleocapsid Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics*; D045473:Severe acute respiratory syndrome-related coronavirus / Q000737:chemistry*; D014763:Viral Matrix Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D000086862:Coronavirus M Proteins; D000086462:Coronavirus Nucleocapsid Proteins; C000706237:M protein, SARS-CoV; D019590:Nucleocapsid Proteins; D012367:RNA, Viral; D014763:Viral Matrix Proteins