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Structural studies of the nucleus-like assembly of jumbo bacteriophage 201φ2-1

The jumbo phages encode proteins that assemble to form a nucleus-like compartment in infected cells. Here we report the cryo-EM structure and biochemistry characterization of gp105, a protein that is encoded by the jumbo phage 201φ2-1 and is involved in the formation of the nucleus-like compartment...

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Autores principales: Liu, Zhe, Xiang, Ye
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10149743/
https://www.ncbi.nlm.nih.gov/pubmed/37138628
http://dx.doi.org/10.3389/fmicb.2023.1170112
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author Liu, Zhe
Xiang, Ye
author_facet Liu, Zhe
Xiang, Ye
author_sort Liu, Zhe
collection PubMed
description The jumbo phages encode proteins that assemble to form a nucleus-like compartment in infected cells. Here we report the cryo-EM structure and biochemistry characterization of gp105, a protein that is encoded by the jumbo phage 201φ2-1 and is involved in the formation of the nucleus-like compartment in phage 201φ2-1 infected Pseudomonas chlororaphis. We found that, although most gp105 molecules are in the monomeric state in solution, a small portion of gp105 assemble to form large sheet-like assemblies and small cube-like particles. Reconstruction of the cube-like particles showed that the particle consists of six flat head-to-tail tetramers arranged into an octahedral cube. The four molecules at the contact interface of two head-to-tail tetramers are 2-fold symmetry-related and constitute a concave tetramer. Further reconstructions without applying symmetry showed that molecules in the particles around the distal ends of a 3-fold axis are highly dynamic and have the tendency to open up the assembly. Local classifications and refinements of the concave tetramers in the cube-like particle resulted in a map of the concave tetramer at a resolution of 4.09 Å. Structural analysis of the concave tetramer indicates that the N and C terminal fragments of gp105 are important for mediating the intermolecular interactions, which was further confirmed by mutagenesis studies. Biochemistry assays showed that, in solution, the cube-like particles of gp105 are liable to either disassemble to form the monomers or recruit more molecules to form the high molecular weight lattice-like assembly. We also found that monomeric gp105s can self-assemble to form large sheet-like assemblies in vitro, and the assembly of gp105 in vitro is a reversible dynamic process and temperature-dependent. Taken together, our results revealed the dynamic assembly of gp105, which helps to understand the development and function of the nucleus-like compartment assembled by phage-encoded proteins.
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spelling pubmed-101497432023-05-02 Structural studies of the nucleus-like assembly of jumbo bacteriophage 201φ2-1 Liu, Zhe Xiang, Ye Front Microbiol Microbiology The jumbo phages encode proteins that assemble to form a nucleus-like compartment in infected cells. Here we report the cryo-EM structure and biochemistry characterization of gp105, a protein that is encoded by the jumbo phage 201φ2-1 and is involved in the formation of the nucleus-like compartment in phage 201φ2-1 infected Pseudomonas chlororaphis. We found that, although most gp105 molecules are in the monomeric state in solution, a small portion of gp105 assemble to form large sheet-like assemblies and small cube-like particles. Reconstruction of the cube-like particles showed that the particle consists of six flat head-to-tail tetramers arranged into an octahedral cube. The four molecules at the contact interface of two head-to-tail tetramers are 2-fold symmetry-related and constitute a concave tetramer. Further reconstructions without applying symmetry showed that molecules in the particles around the distal ends of a 3-fold axis are highly dynamic and have the tendency to open up the assembly. Local classifications and refinements of the concave tetramers in the cube-like particle resulted in a map of the concave tetramer at a resolution of 4.09 Å. Structural analysis of the concave tetramer indicates that the N and C terminal fragments of gp105 are important for mediating the intermolecular interactions, which was further confirmed by mutagenesis studies. Biochemistry assays showed that, in solution, the cube-like particles of gp105 are liable to either disassemble to form the monomers or recruit more molecules to form the high molecular weight lattice-like assembly. We also found that monomeric gp105s can self-assemble to form large sheet-like assemblies in vitro, and the assembly of gp105 in vitro is a reversible dynamic process and temperature-dependent. Taken together, our results revealed the dynamic assembly of gp105, which helps to understand the development and function of the nucleus-like compartment assembled by phage-encoded proteins. Frontiers Media S.A. 2023-04-17 /pmc/articles/PMC10149743/ /pubmed/37138628 http://dx.doi.org/10.3389/fmicb.2023.1170112 Text en Copyright © 2023 Liu and Xiang. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Liu, Zhe
Xiang, Ye
Structural studies of the nucleus-like assembly of jumbo bacteriophage 201φ2-1
title Structural studies of the nucleus-like assembly of jumbo bacteriophage 201φ2-1
title_full Structural studies of the nucleus-like assembly of jumbo bacteriophage 201φ2-1
title_fullStr Structural studies of the nucleus-like assembly of jumbo bacteriophage 201φ2-1
title_full_unstemmed Structural studies of the nucleus-like assembly of jumbo bacteriophage 201φ2-1
title_short Structural studies of the nucleus-like assembly of jumbo bacteriophage 201φ2-1
title_sort structural studies of the nucleus-like assembly of jumbo bacteriophage 201φ2-1
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10149743/
https://www.ncbi.nlm.nih.gov/pubmed/37138628
http://dx.doi.org/10.3389/fmicb.2023.1170112
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