Solving the MCM paradox by visualizing the scaffold of CMG helicase at active replisomes

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Solving the MCM paradox by visualizing the scaffold of CMG helicase at active replisomes. / Polasek-Sedlackova, Hana; Miller, Thomas C.R.; Krejci, Jana; Rask, Maj Britt; Lukas, Jiri.

In: Nature Communications, Vol. 13, 6090, 2022.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Polasek-Sedlackova, H, Miller, TCR, Krejci, J, Rask, MB & Lukas, J 2022, 'Solving the MCM paradox by visualizing the scaffold of CMG helicase at active replisomes', Nature Communications, vol. 13, 6090. https://doi.org/10.1038/s41467-022-33887-5

APA

Polasek-Sedlackova, H., Miller, T. C. R., Krejci, J., Rask, M. B., & Lukas, J. (2022). Solving the MCM paradox by visualizing the scaffold of CMG helicase at active replisomes. Nature Communications, 13, [6090]. https://doi.org/10.1038/s41467-022-33887-5

Vancouver

Polasek-Sedlackova H, Miller TCR, Krejci J, Rask MB, Lukas J. Solving the MCM paradox by visualizing the scaffold of CMG helicase at active replisomes. Nature Communications. 2022;13. 6090. https://doi.org/10.1038/s41467-022-33887-5

Author

Polasek-Sedlackova, Hana ; Miller, Thomas C.R. ; Krejci, Jana ; Rask, Maj Britt ; Lukas, Jiri. / Solving the MCM paradox by visualizing the scaffold of CMG helicase at active replisomes. In: Nature Communications. 2022 ; Vol. 13.

Bibtex

@article{315ec4d847c7443eb4374856c2bdb92f,
title = "Solving the MCM paradox by visualizing the scaffold of CMG helicase at active replisomes",
abstract = "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.",
author = "Hana Polasek-Sedlackova and Miller, {Thomas C.R.} and Jana Krejci and Rask, {Maj Britt} and Jiri Lukas",
note = "Publisher Copyright: {\textcopyright} 2022, The Author(s).",
year = "2022",
doi = "10.1038/s41467-022-33887-5",
language = "English",
volume = "13",
journal = "Nature Communications",
issn = "2041-1723",
publisher = "nature publishing group",

}

RIS

TY - JOUR

T1 - Solving the MCM paradox by visualizing the scaffold of CMG helicase at active replisomes

AU - Polasek-Sedlackova, Hana

AU - Miller, Thomas C.R.

AU - Krejci, Jana

AU - Rask, Maj Britt

AU - Lukas, Jiri

N1 - Publisher Copyright: © 2022, The Author(s).

PY - 2022

Y1 - 2022

N2 - 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.

AB - 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.

U2 - 10.1038/s41467-022-33887-5

DO - 10.1038/s41467-022-33887-5

M3 - Journal article

C2 - 36241664

AN - SCOPUS:85139886579

VL - 13

JO - Nature Communications

JF - Nature Communications

SN - 2041-1723

M1 - 6090

ER -

ID: 323839592