Mechanisms of opening and closing of the bacterial replicative helicase
Assembly of bacterial ring-shaped hexameric replicative helicases on single-stranded (ss) DNA requires specialized loading factors. However, mechanisms implemented by these factors during opening and closing of the helicase, which enable and restrict access to an internal chamber, are not known. Her...
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doaj-c5e7063e0b1b4a42a358ecdbc1d0425f2021-05-05T16:23:08ZengeLife Sciences Publications LtdeLife2050-084X2018-12-01710.7554/eLife.41140Mechanisms of opening and closing of the bacterial replicative helicaseJillian Chase0https://orcid.org/0000-0002-1027-6516Andrew Catalano1Alex J Noble2https://orcid.org/0000-0001-8634-2279Edward T Eng3https://orcid.org/0000-0002-8014-7269Paul DB Olinares4Kelly Molloy5Danaya Pakotiprapha6https://orcid.org/0000-0002-5017-8283Martin Samuels7Brian Chait8Amedee des Georges9David Jeruzalmi10https://orcid.org/0000-0001-5886-1370Department of Chemistry and Biochemistry, City College of New York, New York, United States; PhD Program in Biochemistry, The Graduate Center of the City University of New York, New York, United StatesDepartment of Chemistry and Biochemistry, City College of New York, New York, United StatesSimons Electron Microscopy Center, The New York Structural Biology Center, New York, United StatesSimons Electron Microscopy Center, The New York Structural Biology Center, New York, United StatesLaboratory for Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, United StatesLaboratory for Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, United StatesDepartment of Biochemistry, Center for Excellence in Protein and Enzyme Technology, Faculty of Science, Mahidol University, Bangkok, Thailand; Department of Molecular and Cellular Biology, Harvard University, Cambridge, United StatesDepartment of Molecular and Cellular Biology, Harvard University, Cambridge, United StatesLaboratory for Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, United StatesDepartment of Chemistry and Biochemistry, City College of New York, New York, United States; PhD Program in Biochemistry, The Graduate Center of the City University of New York, New York, United States; Structural Biology Initiative, CUNY Advanced Science Research Center, New York, United States; PhD Program in Chemistry, The Graduate Center of the City University of New York, New York, United StatesDepartment of Chemistry and Biochemistry, City College of New York, New York, United States; PhD Program in Biochemistry, The Graduate Center of the City University of New York, New York, United States; PhD Program in Chemistry, The Graduate Center of the City University of New York, New York, United States; PhD Program in Biology, The Graduate Center of the City University of New York, New York, United StatesAssembly of bacterial ring-shaped hexameric replicative helicases on single-stranded (ss) DNA requires specialized loading factors. However, mechanisms implemented by these factors during opening and closing of the helicase, which enable and restrict access to an internal chamber, are not known. Here, we investigate these mechanisms in the Escherichia coli DnaB helicase•bacteriophage λ helicase loader (λP) complex. We show that five copies of λP bind at DnaB subunit interfaces and reconfigure the helicase into an open spiral conformation that is intermediate to previously observed closed ring and closed spiral forms; reconfiguration also produces openings large enough to admit ssDNA into the inner chamber. The helicase is also observed in a restrained inactive configuration that poises it to close on activating signal, and transition to the translocation state. Our findings provide insights into helicase opening, delivery to the origin and ssDNA entry, and closing in preparation for translocation.https://elifesciences.org/articles/41140DNA replicationDnaB replicative helicasehelicase loaderreplication initiationcryogenic electron microscopystructural biology |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Jillian Chase Andrew Catalano Alex J Noble Edward T Eng Paul DB Olinares Kelly Molloy Danaya Pakotiprapha Martin Samuels Brian Chait Amedee des Georges David Jeruzalmi |
spellingShingle |
Jillian Chase Andrew Catalano Alex J Noble Edward T Eng Paul DB Olinares Kelly Molloy Danaya Pakotiprapha Martin Samuels Brian Chait Amedee des Georges David Jeruzalmi Mechanisms of opening and closing of the bacterial replicative helicase eLife DNA replication DnaB replicative helicase helicase loader replication initiation cryogenic electron microscopy structural biology |
author_facet |
Jillian Chase Andrew Catalano Alex J Noble Edward T Eng Paul DB Olinares Kelly Molloy Danaya Pakotiprapha Martin Samuels Brian Chait Amedee des Georges David Jeruzalmi |
author_sort |
Jillian Chase |
title |
Mechanisms of opening and closing of the bacterial replicative helicase |
title_short |
Mechanisms of opening and closing of the bacterial replicative helicase |
title_full |
Mechanisms of opening and closing of the bacterial replicative helicase |
title_fullStr |
Mechanisms of opening and closing of the bacterial replicative helicase |
title_full_unstemmed |
Mechanisms of opening and closing of the bacterial replicative helicase |
title_sort |
mechanisms of opening and closing of the bacterial replicative helicase |
publisher |
eLife Sciences Publications Ltd |
series |
eLife |
issn |
2050-084X |
publishDate |
2018-12-01 |
description |
Assembly of bacterial ring-shaped hexameric replicative helicases on single-stranded (ss) DNA requires specialized loading factors. However, mechanisms implemented by these factors during opening and closing of the helicase, which enable and restrict access to an internal chamber, are not known. Here, we investigate these mechanisms in the Escherichia coli DnaB helicase•bacteriophage λ helicase loader (λP) complex. We show that five copies of λP bind at DnaB subunit interfaces and reconfigure the helicase into an open spiral conformation that is intermediate to previously observed closed ring and closed spiral forms; reconfiguration also produces openings large enough to admit ssDNA into the inner chamber. The helicase is also observed in a restrained inactive configuration that poises it to close on activating signal, and transition to the translocation state. Our findings provide insights into helicase opening, delivery to the origin and ssDNA entry, and closing in preparation for translocation. |
topic |
DNA replication DnaB replicative helicase helicase loader replication initiation cryogenic electron microscopy structural biology |
url |
https://elifesciences.org/articles/41140 |
work_keys_str_mv |
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