title: Crystal structure of the C-terminal domain of Mu phage central spike and functions of bound calcium ion.
abstract: Bacteriophage Mu, which has a contractile tail, is one of the most famous genus of Myoviridae. It has a wide host range and is thought to contribute to horizontal gene transfer. The Myoviridae infection process is initiated by adhesion to the host surface. The phage then penetrates the host cell membrane using its tail to inject its genetic material into the host. In this penetration process, Myoviridae phages are proposed to puncture the membrane of the host cell using a central spike located beneath its baseplate. The central spike of the Mu phage is thought to be composed of gene 45 product (gp45), which has a significant sequence homology with the central spike of P2 phage (gpV). We determined the crystal structure of shortened Mu gp45Δ1-91 (Arg92-Gln197) at 1.5Å resolution and showed that Mu gp45 is a needlelike structure that punctures the membrane. The apex of Mu gp45 and that of P2 gpV contained iron, chloride, and calcium ions. Although the C-terminal domain of Mu gp45 was sufficient for binding to the E. coli membrane, a mutant D188A, in which the Asp amino acid residue that coordinates the calcium ion was replaced by Ala, did not exhibit a propensity to bind to the membrane. Therefore, we concluded that calcium ion played an important role in interaction with the host cell membrane.
authors.affiliation: Institute for Protein Research, Osaka University, 3-2 Yamada-oka, Suita, Osaka 565-0871, Japan.
authors.forename: Kenichi
authors.initials: K
authors.lastname: Harada
authors.forename: Eiki
authors.initials: E
authors.lastname: Yamashita
authors.forename: Atsushi
authors.initials: A
authors.lastname: Nakagawa
authors.forename: Takamitsu
authors.initials: T
authors.lastname: Miyafusa
authors.forename: Kouhei
authors.initials: K
authors.lastname: Tsumoto
authors.forename: Takashi
authors.initials: T
authors.lastname: Ueno
authors.forename: Yoshiharu
authors.initials: Y
authors.lastname: Toyama
authors.forename: Shigeki
authors.initials: S
authors.lastname: Takeda
mesh_terms: D010583:Bacteriophage mu / Q000737:chemistry* / Q000378:metabolism; D002118:Calcium / Q000737:chemistry / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism / Q000821:virology; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000737:chemistry / Q000378:metabolism / Q000821:virology; D017434:Protein Structure, Tertiary; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000378:metabolism; D053586:Virus Internalization
chemical_list: D014759:Viral Envelope Proteins; D002118:Calcium