Solving the MCM paradox by visualizing the scaffold of CMG helicase at active replisomes
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68081707%3A_____%2F22%3A00564037" target="_blank" >RIV/68081707:_____/22:00564037 - isvavai.cz</a>
Result on the web
<a href="https://www.nature.com/articles/s41467-022-33887-5" target="_blank" >https://www.nature.com/articles/s41467-022-33887-5</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1038/s41467-022-33887-5" target="_blank" >10.1038/s41467-022-33887-5</a>
Alternative languages
Result language
angličtina
Original language name
Solving the MCM paradox by visualizing the scaffold of CMG helicase at active replisomes
Original language description
Genome duplication is safeguarded by constantly adjusting the activity of the replicative CMG (CDC45-MCM2-7-GINS) helicase. However, minichromosome maintenance proteins (MCMs)-the structural core of the CMG helicase-have never been visualized at sites of DNA synthesis inside a cell (the so-called MCM paradox). Here, we solve this conundrum by showing that anti-MCM antibodies primarily detect inactive MCMs. Upon conversion of inactive MCMs to CMGs, factors that are required for replisome activity bind to the MCM scaffold and block MCM antibody binding sites. Tagging of endogenous MCMs by CRISPR-Cas9 bypasses this steric hindrance and enables MCM visualization at active replisomes. Thus, by defining conditions for detecting the structural core of the replicative CMG helicase, our results explain the MCM paradox, provide visual proof that MCMs are an integral part of active replisomes in vivo, and enable the investigation of replication dynamics in living cells exposed to a constantly changing environment.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
—
OECD FORD branch
10601 - Cell biology
Result continuities
Project
<a href="/en/project/GM22-20303M" target="_blank" >GM22-20303M: Deciphering Origins of DNA replication in genome integrity</a><br>
Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2022
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Nature Communications
ISSN
2041-1723
e-ISSN
2041-1723
Volume of the periodical
13
Issue of the periodical within the volume
1
Country of publishing house
GB - UNITED KINGDOM
Number of pages
11
Pages from-to
6090
UT code for WoS article
000874078500024
EID of the result in the Scopus database
2-s2.0-85139886579