In silico reconstitution of DNA replication: Lessons from single-molecule imaging and cryo-tomography applied to single-particle cryo-EM

Julia F. Greiwe; Giulia Zanetti; Thomas C.R. Miller; Alessandro Costa

doi:10.1016/j.sbi.2021.11.015

In silico reconstitution of DNA replication: Lessons from single-molecule imaging and cryo-tomography applied to single-particle cryo-EM

Julia F. Greiwe, Giulia Zanetti, Thomas C.R. Miller, Alessandro Costa^*

^*Corresponding author af dette arbejde

Molecular Aging Program

Publikation: Bidrag til tidsskrift › Review › peer review

3 Citationer (Scopus)

12 Downloads (Pure)

Abstract

DNA replication has been reconstituted in vitro with yeast proteins, and the minimal system requires the coordinated assembly of 16 distinct replication factors, consisting of 42 polypeptides. To understand the molecular interplay between these factors at the single residue level, new structural biology tools are being developed. Inspired by advances in single-molecule fluorescence imaging and cryo-tomography, novel single-particle cryo-EM experiments have been used to characterise the structural mechanism for the loading of the replicative helicase. Here, we discuss how in silico reconstitution of single-particle cryo-EM data can help describe dynamic systems that are difficult to approach with conventional three-dimensional classification tools.

Originalsprog	Engelsk
Tidsskrift	Current Opinion in Structural Biology
Vol/bind	72
Sider (fra-til)	279-286
Antal sider	8
ISSN	0959-440X
DOI	https://doi.org/10.1016/j.sbi.2021.11.015
Status	Udgivet - 2022

Bibliografisk note

Funding Information:
The authors would like to thank the members of the Costa and Zanetti laboratories for helpful discussion. Work in A.C.?s laboratory is funded jointly by the Wellcome Trust, MRC and CRUK at the Francis Crick Institute (FC001065) and by the European Research Council under the European Union's Horizon 2020 research and innovation programme (grant agreement no. 820102). Work in G.Z.?s laboratory is funded by BBSRC (BB/T002670/1) and by the (ERC) under the European Union's Horizon 2020 research and innovation programme (grant agreement StG 852915 ? CRYTOCOP).

Funding Information:
The authors would like to thank the members of the Costa and Zanetti laboratories for helpful discussion. Work in A.C.’s laboratory is funded jointly by the Wellcome Trust , MRC and CRUK at the Francis Crick Institute ( FC001065 ) and by the European Research Council under the European Union’s Horizon 2020 research and innovation programme (grant agreement no. 820102 ). Work in G.Z.’s laboratory is funded by BBSRC ( BB/T002670/1 ) and by the (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement StG 852915 – CRYTOCOP ).

Publisher Copyright:
© 2022 The Authors

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Citationsformater

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abstract = "DNA replication has been reconstituted in vitro with yeast proteins, and the minimal system requires the coordinated assembly of 16 distinct replication factors, consisting of 42 polypeptides. To understand the molecular interplay between these factors at the single residue level, new structural biology tools are being developed. Inspired by advances in single-molecule fluorescence imaging and cryo-tomography, novel single-particle cryo-EM experiments have been used to characterise the structural mechanism for the loading of the replicative helicase. Here, we discuss how in silico reconstitution of single-particle cryo-EM data can help describe dynamic systems that are difficult to approach with conventional three-dimensional classification tools.",

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T2 - Lessons from single-molecule imaging and cryo-tomography applied to single-particle cryo-EM

AU - Greiwe, Julia F.

AU - Zanetti, Giulia

AU - Miller, Thomas C.R.

AU - Costa, Alessandro

PY - 2022

Y1 - 2022

N2 - DNA replication has been reconstituted in vitro with yeast proteins, and the minimal system requires the coordinated assembly of 16 distinct replication factors, consisting of 42 polypeptides. To understand the molecular interplay between these factors at the single residue level, new structural biology tools are being developed. Inspired by advances in single-molecule fluorescence imaging and cryo-tomography, novel single-particle cryo-EM experiments have been used to characterise the structural mechanism for the loading of the replicative helicase. Here, we discuss how in silico reconstitution of single-particle cryo-EM data can help describe dynamic systems that are difficult to approach with conventional three-dimensional classification tools.

AB - DNA replication has been reconstituted in vitro with yeast proteins, and the minimal system requires the coordinated assembly of 16 distinct replication factors, consisting of 42 polypeptides. To understand the molecular interplay between these factors at the single residue level, new structural biology tools are being developed. Inspired by advances in single-molecule fluorescence imaging and cryo-tomography, novel single-particle cryo-EM experiments have been used to characterise the structural mechanism for the loading of the replicative helicase. Here, we discuss how in silico reconstitution of single-particle cryo-EM data can help describe dynamic systems that are difficult to approach with conventional three-dimensional classification tools.

U2 - 10.1016/j.sbi.2021.11.015

DO - 10.1016/j.sbi.2021.11.015

M3 - Review

C2 - 35026552

AN - SCOPUS:85122526109

SN - 0959-440X

VL - 72

SP - 279

EP - 286

JO - Current Opinion in Structural Biology

JF - Current Opinion in Structural Biology

ER -