Zygotic genome activation triggers the DNA replication checkpoint at the midblastula transition

Shelby A. Blythe; Eric F. Wieschaus

doi:10.1016/j.cell.2015.01.050

Zygotic genome activation triggers the DNA replication checkpoint at the midblastula transition

Shelby A. Blythe, Eric F. Wieschaus^*

^*Corresponding author for this work

Molecular Biosciences

Research output: Contribution to journal › Article › peer-review

134 Scopus citations

Abstract

A conserved feature of the midblastula transition (MBT) is a requirement for a functional DNA replication checkpoint to coordinate cell-cycle remodeling and zygotic genome activation (ZGA). We have investigated what triggers this checkpoint during Drosophila embryogenesis. We find that the magnitude of the checkpoint scales with the quantity of transcriptionally engaged DNA. Measuring RNA polymerase II (Pol II) binding at 20 min intervals over the course of ZGA reveals that the checkpoint coincides with widespread de novo recruitment of Pol II that precedes and does not require a functional checkpoint. This recruitment drives slowing or stalling of DNA replication at transcriptionally engaged loci. Reducing Pol II recruitment in zelda mutants both reduces replication stalling and bypasses the requirement for a functional checkpoint. This suggests a model where the checkpoint functions as a feedback mechanism to remodel the cell cycle in response to nascent ZGA.

Original language	English (US)
Pages (from-to)	1169-1181
Number of pages	13
Journal	Cell
Volume	160
Issue number	6
DOIs	https://doi.org/10.1016/j.cell.2015.01.050
State	Published - Mar 15 2015

Funding

We thank the Bloomington Drosophila Stock Center for fly stocks. We thank C. Hannon, A. Roknabadi, and B. Pelham-Webb for help with experiments, all members of the E.F.W. and Schupbach laboratories, especially S. Little and B. He for lively discussion. We thank R. Kadzik, P. Klein, T. Schupbach, J. Lieb, M. Harrison, J. Zeitlinger, T. Orr-Weaver, W. Sullivan, and P. O’Farrell for helpful comments and C. Rushlow, P. Schedl, and J. Sekelsky for fly stocks. We are indebted to the staff of the Sequencing Core Facility of the Lewis Sigler Institute and to L. Parsons for support for the ChIP-seq studies. We thank S. Di Talia for comments on the manuscript. This work was supported in part by grant 5R37HD15587 from the National Institute of Child Health and Human Development (NICHD) to E.F.W. and Ruth Kirschstein National Research Service Award (NRSA) Postdoctoral Fellowship 1F32HD072653 from NICHD to S.A.B. E.F.W. is an investigator with the Howard Hughes Medical Institute.

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology

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10.1016/j.cell.2015.01.050

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title = "Zygotic genome activation triggers the DNA replication checkpoint at the midblastula transition",

abstract = "A conserved feature of the midblastula transition (MBT) is a requirement for a functional DNA replication checkpoint to coordinate cell-cycle remodeling and zygotic genome activation (ZGA). We have investigated what triggers this checkpoint during Drosophila embryogenesis. We find that the magnitude of the checkpoint scales with the quantity of transcriptionally engaged DNA. Measuring RNA polymerase II (Pol II) binding at 20 min intervals over the course of ZGA reveals that the checkpoint coincides with widespread de novo recruitment of Pol II that precedes and does not require a functional checkpoint. This recruitment drives slowing or stalling of DNA replication at transcriptionally engaged loci. Reducing Pol II recruitment in zelda mutants both reduces replication stalling and bypasses the requirement for a functional checkpoint. This suggests a model where the checkpoint functions as a feedback mechanism to remodel the cell cycle in response to nascent ZGA.",

author = "Blythe, {Shelby A.} and Wieschaus, {Eric F.}",

note = "Funding Information: We thank the Bloomington Drosophila Stock Center for fly stocks. We thank C. Hannon, A. Roknabadi, and B. Pelham-Webb for help with experiments, all members of the E.F.W. and Schupbach laboratories, especially S. Little and B. He for lively discussion. We thank R. Kadzik, P. Klein, T. Schupbach, J. Lieb, M. Harrison, J. Zeitlinger, T. Orr-Weaver, W. Sullivan, and P. O{\textquoteright}Farrell for helpful comments and C. Rushlow, P. Schedl, and J. Sekelsky for fly stocks. We are indebted to the staff of the Sequencing Core Facility of the Lewis Sigler Institute and to L. Parsons for support for the ChIP-seq studies. We thank S. Di Talia for comments on the manuscript. This work was supported in part by grant 5R37HD15587 from the National Institute of Child Health and Human Development (NICHD) to E.F.W. and Ruth Kirschstein National Research Service Award (NRSA) Postdoctoral Fellowship 1F32HD072653 from NICHD to S.A.B. E.F.W. is an investigator with the Howard Hughes Medical Institute. Publisher Copyright: {\textcopyright} 2015 Elsevier Inc.",

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Zygotic genome activation triggers the DNA replication checkpoint at the midblastula transition

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