scholarly journals High Resolution Structure of the Mature Capsid of Ralstonia Solanacearum Bacteriophage ϕRSA1 by Cryo-Electron Microscopy

2021 ◽  
Vol 22 (20) ◽  
pp. 11053
Author(s):  
Grégory Effantin ◽  
Akiko Fujiwara ◽  
Takeru Kawasaki ◽  
Takashi Yamada ◽  
Guy Schoehn
Keyword(s):  
Electron Microscopy ◽  
Ralstonia Solanacearum ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Icosahedral Symmetry ◽  
E Coli ◽  
Covalent Crosslinking ◽  
Cryo Electron Microscopy ◽  
Capsid Shell ◽  
High Resolution Structure

The ϕRSA1 bacteriophage has been isolated from Ralstonia solanacearum, a gram negative bacteria having a significant economic impact on many important crops. We solved the three-dimensional structure of the ϕRSA1 mature capsid to 3.9 Å resolution by cryo-electron microscopy. The capsid shell, that contains the 39 kbp of dsDNA genome, has an icosahedral symmetry characterized by an unusual triangulation number of T = 7, dextro. The ϕRSA1 capsid is composed solely of the polymerization of the major capsid protein, gp8, which exhibits the typical “Johnson” fold first characterized in E. coli bacteriophage HK97. As opposed to the latter, the ϕRSA1 mature capsid is not stabilized by covalent crosslinking between its subunits, nor by the addition of a decoration protein. We further describe the molecular interactions occurring between the subunits of the ϕRSA1 capsid and their relationships with the other known bacteriophages.

scholarly journals Cryo-Electron Microscopy Three-Dimensional Structure of the Jumbo Phage ΦRSL1 Infecting the Phytopathogen Ralstonia solanacearum

Structure ◽  
2013 ◽  
Vol 21 (2) ◽  
pp. 298-305 ◽  
Author(s):  
Grégory Effantin ◽  
Ryosuke Hamasaki ◽  
Takeru Kawasaki ◽  
Maria Bacia ◽  
Christine Moriscot ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Ralstonia Solanacearum ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Three Dimensional Structure ◽  
Cryo Electron Microscopy

scholarly journals Capturing Enveloped Viruses on Affinity Grids for Downstream Cryo-Electron Microscopy Applications

2013 ◽  
Vol 20 (1) ◽  
pp. 164-174 ◽  
Author(s):  
Gabriella Kiss ◽  
Xuemin Chen ◽  
Melinda A. Brindley ◽  
Patricia Campbell ◽  
Claudio L. Afonso ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Measles Virus ◽  
Influenza A ◽  
Electron Tomography ◽  
Three Dimensional ◽  
Nitrilotriacetic Acid ◽  
Dimensional Structure ◽  
Enveloped Viruses ◽  
Cryo Electron Microscopy ◽  
Human Immunodeficiency Virus Virus

AbstractElectron microscopy (EM), cryo-electron microscopy (cryo-EM), and cryo-electron tomography (cryo-ET) are essential techniques used for characterizing basic virus morphology and determining the three-dimensional structure of viruses. Enveloped viruses, which contain an outer lipoprotein coat, constitute the largest group of pathogenic viruses to humans. The purification of enveloped viruses from cell culture presents certain challenges. Specifically, the inclusion of host-membrane-derived vesicles, the complete destruction of the viruses, and the disruption of the internal architecture of individual virus particles. Here, we present a strategy for capturing enveloped viruses on affinity grids (AG) for use in both conventional EM and cryo-EM/ET applications. We examined the utility of AG for the selective capture of human immunodeficiency virus virus-like particles, influenza A, and measles virus. We applied nickel-nitrilotriacetic acid lipid layers in combination with molecular adaptors to selectively adhere the viruses to the AG surface. This further development of the AG method may prove essential for the gentle and selective purification of enveloped viruses directly onto EM grids for ultrastructural analyses.

The Ribosome - Three-Dimensional Structure and Ligand-Binding Studies

1998 ◽  
Vol 4 (S2) ◽  
pp. 962-963
Author(s):  
J. Frank ◽  
P. Penczek ◽  
A. Malhotra ◽  
I. Gabashvili ◽  
R. Grassucci ◽  
...  
Keyword(s):  
State Of The Art ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Conformational Heterogeneity ◽  
Binding Studies ◽  
E Coli ◽  
Cryo Electron Microscopy ◽  
Protein Synthesis Initiation ◽  
Successful Technique ◽  
Electron Microscopes

To date, cryo-electron microscopy has become the most successful technique for exploring the structure of the ribosome and for studying binding positions of its various ligands, with the resolution slowly extending toward 10 Å. Obstacles in the attempts to improve resolution are the limited stability and coherence of the electron microscope, the statistics of data collection, and the conformational heterogeneity of the specimen. The last factor in this list proved to be the reason why it has been difficult to go past 18-20 Å with many specimens despite the use of state-of-the-art electron microscopes and inclusion of tens of thousand of projections.A breakthrough has been achieved with a protein synthesis initiation-like complex in which mRNA and fMet-tRNA is bound to the E. coli ribosome. The high occupancy and extraordinary conformational homogeneity of this specimen has enabled us to reach a resolution of 15 Å.

scholarly journals 3P131 The three-dimensional structure of FtsH oligomer from E. coli by cryonegative electron-microscopy

2005 ◽  
Vol 45 (supplement) ◽  
pp. S236
Author(s):  
N. Saikawa ◽  
H. Suzuki ◽  
Y. Akiyama ◽  
K. Ito ◽  
Y. Kimura
Keyword(s):  
Electron Microscopy ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Three Dimensional Structure ◽  
E Coli
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