Metabolic remodeling maintains a reducing environment for rapid activation of the yeast DNA replication checkpoint

Abstract

Nucleotide metabolism fuels normal DNA replication and is also primarily targeted by the DNA replication checkpoint when replication stalls. To reveal a comprehensive interconnection between genome maintenance and metabolism, we analyzed the metabolomic changes upon replication stress in the budding yeast S. cerevisiae. We found that upon treatment of cells with hydroxyurea, glucose is rapidly diverted to the oxidative pentose phosphate pathway (PPP). This effect is mediated by the AMP-dependent kinase, SNF1, which phosphorylates the transcription factor Mig1, thereby relieving repression of the gene encoding the rate-limiting enzyme of the PPP. Surprisingly, NADPH produced by the PPP is required for efficient recruitment of replication protein A (RPA) to single-stranded DNA, providing the signal for the activation of the Mec1/ATR-Rad53/CHK1 checkpoint signaling kinase cascade. Thus, SNF1, best known as a central energy controller, determines a fast mode of replication checkpoint activation through a redox mechanism. These findings establish that SNF1 provides a hub with direct links to cellular metabolism, redox, and surveillance of DNA replication in eukaryotes.

Addresses

State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China.

Autoren

Lili Li
Jie Wang
Zijia Yang
Yiling Zhao
Hui Jiang
Luguang Jiang
Wenya Hou
Risheng Ye
Qun He
Martin Kupiec
Brian Luke
Qinhong Cao
Zhi Qi
Zhen Li
Huiqiang Lou

DOI

10.15252/embj.2021108290

eISSN

1460-2075

Externe Identifier

PubMed Identifier: 35028974
PubMed Central ID: PMC8844976

Funding acknowledgements

Foundation for Innovative Research Groups of the National Natural Science Foundation of China: 31670762
Foundation for Innovative Research Groups of the National Natural Science Foundation of China: 32161133015
Foundation for Innovative Research Groups of the National Natural Science Foundation of China: 31770084
National Key Research and Development Program of China: 2019YFA0903900
Foundation for Innovative Research Groups of the National Natural Science Foundation of China: 31771382
Foundation for Innovative Research Groups of the National Natural Science Foundation of China: 31630005
Foundation for Innovative Research Groups of the National Natural Science Foundation of China: 32101039

Open access

false

ISSN

0261-4189

Ausgabe der Veröffentlichung

4

Zeitschrift

The EMBO journal

Schlüsselwörter

Saccharomyces cerevisiae
Hydroxyurea
NADP
Glucose
Intracellular Signaling Peptides and Proteins
Cell Cycle Proteins
Saccharomyces cerevisiae Proteins
Repressor Proteins
DNA, Single-Stranded
DNA Replication
Glycolysis
Phosphorylation
Pentose Phosphate Pathway
Replication Protein A
Checkpoint Kinase 2
Protein Serine-Threonine Kinases

Sprache

eng

Medium

Print-Electronic

Online publication date

2022

Paginierung

e108290

Datum der Veröffentlichung

2022

Status

Published

Datum der Datenerfassung

2022

Titel

Metabolic remodeling maintains a reducing environment for rapid activation of the yeast DNA replication checkpoint.

Sub types

Research Support, Non-U.S. Gov't
research-article
Journal Article

Ausgabe der Zeitschrift

41

Files

https://europepmc.org/articles/PMC8844976?pdf=render

Datenquelle: Europe PubMed Central

Abstract

Nucleotide metabolism fuels normal DNA replication and is also primarily targeted by the DNA replication checkpoint when replication stalls. To reveal a comprehensive interconnection between genome maintenance and metabolism, we analyzed the metabolomic changes upon replication stress in the budding yeast S. cerevisiae. We found that upon treatment of cells with hydroxyurea, glucose is rapidly diverted to the oxidative pentose phosphate pathway (PPP). This effect is mediated by the AMP-dependent kinase, SNF1, which phosphorylates the transcription factor Mig1, thereby relieving repression of the gene encoding the rate-limiting enzyme of the PPP. Surprisingly, NADPH produced by the PPP is required for efficient recruitment of replication protein A (RPA) to single-stranded DNA, providing the signal for the activation of the Mec1/ATR-Rad53/CHK1 checkpoint signaling kinase cascade. Thus, SNF1, best known as a central energy controller, determines a fast mode of replication checkpoint activation through a redox mechanism. These findings establish that SNF1 provides a hub with direct links to cellular metabolism, redox, and surveillance of DNA replication in eukaryotes.

Date of acceptance

2021

Autoren

Lili Li
Jie Wang
Zijia Yang
Yiling Zhao
Hui Jiang
Luguang Jiang
Wenya Hou
Risheng Ye
Qun He
Martin Kupiec
Brian Luke
Qinhong Cao
Zhi Qi
Zhen Li
Huiqiang Lou

Autoren-URL

https://www.ncbi.nlm.nih.gov/pubmed/35028974

DOI

10.15252/embj.2021108290

eISSN

1460-2075

Externe Identifier

PubMed Central ID: PMC8844976

Ausgabe der Veröffentlichung

4

Zeitschrift

EMBO J

Schlüsselwörter

DNA replication stress
carbon metabolism
cell cycle checkpoints
genome stability
reductive/oxidative (redox)
Cell Cycle Proteins
Checkpoint Kinase 2
DNA Replication
DNA, Single-Stranded
Glucose
Glycolysis
Hydroxyurea
Intracellular Signaling Peptides and Proteins
NADP
Pentose Phosphate Pathway
Phosphorylation
Protein Serine-Threonine Kinases
Replication Protein A
Repressor Proteins
Saccharomyces cerevisiae
Saccharomyces cerevisiae Proteins

Sprache

eng

Country

England

Paginierung

e108290

Datum der Veröffentlichung

2022

Status

Published

Datum, an dem der Datensatz öffentlich gemacht wurde

2022

Titel

Metabolic remodeling maintains a reducing environment for rapid activation of the yeast DNA replication checkpoint.

Sub types

Journal Article
Research Support, Non-U.S. Gov't

Ausgabe der Zeitschrift

41

Datenquelle: PubMed

Abstract

Nucleotide metabolism fuels normal DNA replication and is also primarily targeted by the DNA replication checkpoint when replication stalls. To reveal a comprehensive interconnection between genome maintenance and metabolism, we analyzed the metabolomic changes upon replication stress in the budding yeast S. cerevisiae. We found that upon treatment of cells with hydroxyurea, glucose is rapidly diverted to the oxidative pentose phosphate pathway (PPP). This effect is mediated by the AMP-dependent kinase, SNF1, which phosphorylates the transcription factor Mig1, thereby relieving repression of the gene encoding the rate-limiting enzyme of the PPP. Surprisingly, NADPH produced by the PPP is required for efficient recruitment of replication protein A (RPA) to single-stranded DNA, providing the signal for the activation of the Mec1/ATR-Rad53/CHK1 checkpoint signaling kinase cascade. Thus, SNF1, best known as a central energy controller, determines a fast mode of replication checkpoint activation through a redox mechanism. These findings establish that SNF1 provides a hub with direct links to cellular metabolism, redox, and surveillance of DNA replication in eukaryotes.

Autoren

Lili Li
Jie Wang
Zijia Yang
Yiling Zhao
Hui Jiang
Luguang Jiang
Wenya Hou
Risheng Ye
Qun He
Martin Kupiec
Brian Luke
Qinhong Cao
Zhi Qi
Zhen Li
Huiqiang Lou

DOI

10.15252/embj.2021108290

Zeitschrift

The EMBO journal

Notes

keywords: Cell Cycle Proteins/genetics/metabolism;Checkpoint Kinase 2/genetics/metabolism;DNA Replication/drug effects;DNA, Single-Stranded/metabolism;Glucose/genetics/metabolism;Glycolysis/physiology;Hydroxyurea;Intracellular Signaling Peptides and Proteins/genetics/metabolism;NADP/metabolism;Pentose Phosphate Pathway;Phosphorylation;Protein Serine-Threonine Kinases/genetics/metabolism;Replication Protein A/genetics/metabolism;Repressor Proteins/genetics/metabolism;Saccharomyces cerevisiae Proteins/genetics/metabolism;Saccharomyces cerevisiae/drug effects/genetics/metabolism

Artikelnummer

4

Paginierung

e108290 - e108290

Datum der Veröffentlichung

2022

Datum der Datenerfassung

2023

Titel

Metabolic remodeling maintains a reducing environment for rapid activation of the yeast DNA replication checkpoint

Sub types

article

Ausgabe der Zeitschrift

41

Datenquelle: Manual

Metabolic remodeling maintains a reducing environment for rapid activation of the yeast DNA replication checkpoint

Files

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