Electron microscopic imaging revealed the flexible filamentous structure of the cell attachment protein P2 of Rice dwarf virus located around the icosahedral 5-fold axes

Naoyuki Miyazaki, Akifumi Higashiura, Tomoko Higashiura, Fusamichi Akita, Hiroyuki Hibino, Toshihiro Omura, Atsushi Nakagawa, Kenji Iwasaki

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

The minor outer capsid protein P2 of Rice dwarf virus (RDV), a member of the genus Phytoreovirus in the family Reoviridae, is essential for viral cell entry. Here, we clarified the structure of P2 and the interactions to host insect cells. Negative stain electron microscopy (EM) showed that P2 proteins are monomeric and flexible L-shaped filamentous structures of ∼20 nm in length. Cryo-EM structure revealed the spatial arrangement of P2 in the capsid, which was prescribed by the characteristic virion structure. The P2 proteins were visualized as partial rod-shaped structures of ∼10 nm in length in the cryo-EM map and accommodated in crevasses on the viral surface around icosahedral 5-fold axes with hydrophobic interactions. The remaining disordered region of P2 assumed to be extended to the radial direction towards exterior. Electron tomography clearly showed that RDV particles were away from the cellular membrane at a uniform distance and several spike-like densities, probably corresponding to P2, connecting a viral particle to the host cellular membrane during cell entry. By combining the in vitro and in vivo structural information, we could gain new insights into the detailed mechanism of the cell entry of RDV.

Original languageEnglish
Pages (from-to)181-190
Number of pages10
JournalJournal of Biochemistry
Volume159
Issue number2
DOIs
Publication statusPublished - May 28 2015

Fingerprint

Flexible structures
Reoviridae
Viruses
Virion
Electron microscopy
Cryoelectron Microscopy
Electrons
Imaging techniques
Electron Microscope Tomography
Proteins
Capsid
Capsid Proteins
Cell membranes
Hydrophobic and Hydrophilic Interactions
Tomography
Insects
Electron Microscopy
Coloring Agents
Cell Membrane
Membranes

Keywords

  • cryo-electron microscopy
  • electron tomography
  • Phytoreovirus
  • Rice dwarf virus
  • viral cell entry

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Electron microscopic imaging revealed the flexible filamentous structure of the cell attachment protein P2 of Rice dwarf virus located around the icosahedral 5-fold axes. / Miyazaki, Naoyuki; Higashiura, Akifumi; Higashiura, Tomoko; Akita, Fusamichi; Hibino, Hiroyuki; Omura, Toshihiro; Nakagawa, Atsushi; Iwasaki, Kenji.

In: Journal of Biochemistry, Vol. 159, No. 2, 28.05.2015, p. 181-190.

Research output: Contribution to journalArticle

Miyazaki, Naoyuki ; Higashiura, Akifumi ; Higashiura, Tomoko ; Akita, Fusamichi ; Hibino, Hiroyuki ; Omura, Toshihiro ; Nakagawa, Atsushi ; Iwasaki, Kenji. / Electron microscopic imaging revealed the flexible filamentous structure of the cell attachment protein P2 of Rice dwarf virus located around the icosahedral 5-fold axes. In: Journal of Biochemistry. 2015 ; Vol. 159, No. 2. pp. 181-190.
@article{4d7a90ecccd441c6bd6b1d4a57a82db6,
title = "Electron microscopic imaging revealed the flexible filamentous structure of the cell attachment protein P2 of Rice dwarf virus located around the icosahedral 5-fold axes",
abstract = "The minor outer capsid protein P2 of Rice dwarf virus (RDV), a member of the genus Phytoreovirus in the family Reoviridae, is essential for viral cell entry. Here, we clarified the structure of P2 and the interactions to host insect cells. Negative stain electron microscopy (EM) showed that P2 proteins are monomeric and flexible L-shaped filamentous structures of ∼20 nm in length. Cryo-EM structure revealed the spatial arrangement of P2 in the capsid, which was prescribed by the characteristic virion structure. The P2 proteins were visualized as partial rod-shaped structures of ∼10 nm in length in the cryo-EM map and accommodated in crevasses on the viral surface around icosahedral 5-fold axes with hydrophobic interactions. The remaining disordered region of P2 assumed to be extended to the radial direction towards exterior. Electron tomography clearly showed that RDV particles were away from the cellular membrane at a uniform distance and several spike-like densities, probably corresponding to P2, connecting a viral particle to the host cellular membrane during cell entry. By combining the in vitro and in vivo structural information, we could gain new insights into the detailed mechanism of the cell entry of RDV.",
keywords = "cryo-electron microscopy, electron tomography, Phytoreovirus, Rice dwarf virus, viral cell entry",
author = "Naoyuki Miyazaki and Akifumi Higashiura and Tomoko Higashiura and Fusamichi Akita and Hiroyuki Hibino and Toshihiro Omura and Atsushi Nakagawa and Kenji Iwasaki",
year = "2015",
month = "5",
day = "28",
doi = "10.1093/jb/mvv092",
language = "English",
volume = "159",
pages = "181--190",
journal = "Journal of Biochemistry",
issn = "0021-924X",
publisher = "Oxford University Press",
number = "2",

}

TY - JOUR

T1 - Electron microscopic imaging revealed the flexible filamentous structure of the cell attachment protein P2 of Rice dwarf virus located around the icosahedral 5-fold axes

AU - Miyazaki, Naoyuki

AU - Higashiura, Akifumi

AU - Higashiura, Tomoko

AU - Akita, Fusamichi

AU - Hibino, Hiroyuki

AU - Omura, Toshihiro

AU - Nakagawa, Atsushi

AU - Iwasaki, Kenji

PY - 2015/5/28

Y1 - 2015/5/28

N2 - The minor outer capsid protein P2 of Rice dwarf virus (RDV), a member of the genus Phytoreovirus in the family Reoviridae, is essential for viral cell entry. Here, we clarified the structure of P2 and the interactions to host insect cells. Negative stain electron microscopy (EM) showed that P2 proteins are monomeric and flexible L-shaped filamentous structures of ∼20 nm in length. Cryo-EM structure revealed the spatial arrangement of P2 in the capsid, which was prescribed by the characteristic virion structure. The P2 proteins were visualized as partial rod-shaped structures of ∼10 nm in length in the cryo-EM map and accommodated in crevasses on the viral surface around icosahedral 5-fold axes with hydrophobic interactions. The remaining disordered region of P2 assumed to be extended to the radial direction towards exterior. Electron tomography clearly showed that RDV particles were away from the cellular membrane at a uniform distance and several spike-like densities, probably corresponding to P2, connecting a viral particle to the host cellular membrane during cell entry. By combining the in vitro and in vivo structural information, we could gain new insights into the detailed mechanism of the cell entry of RDV.

AB - The minor outer capsid protein P2 of Rice dwarf virus (RDV), a member of the genus Phytoreovirus in the family Reoviridae, is essential for viral cell entry. Here, we clarified the structure of P2 and the interactions to host insect cells. Negative stain electron microscopy (EM) showed that P2 proteins are monomeric and flexible L-shaped filamentous structures of ∼20 nm in length. Cryo-EM structure revealed the spatial arrangement of P2 in the capsid, which was prescribed by the characteristic virion structure. The P2 proteins were visualized as partial rod-shaped structures of ∼10 nm in length in the cryo-EM map and accommodated in crevasses on the viral surface around icosahedral 5-fold axes with hydrophobic interactions. The remaining disordered region of P2 assumed to be extended to the radial direction towards exterior. Electron tomography clearly showed that RDV particles were away from the cellular membrane at a uniform distance and several spike-like densities, probably corresponding to P2, connecting a viral particle to the host cellular membrane during cell entry. By combining the in vitro and in vivo structural information, we could gain new insights into the detailed mechanism of the cell entry of RDV.

KW - cryo-electron microscopy

KW - electron tomography

KW - Phytoreovirus

KW - Rice dwarf virus

KW - viral cell entry

UR - http://www.scopus.com/inward/record.url?scp=84960108654&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84960108654&partnerID=8YFLogxK

U2 - 10.1093/jb/mvv092

DO - 10.1093/jb/mvv092

M3 - Article

VL - 159

SP - 181

EP - 190

JO - Journal of Biochemistry

JF - Journal of Biochemistry

SN - 0021-924X

IS - 2

ER -