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Rad53 arrests leading and lagging strand DNA synthesis via distinct mechanisms in response to DNA replication stress

Richard He1,2,3,4, Zhiguo Zhang1,2,3,4

  • 1Institute for Cancer Genetics, Columbia University Irving Medical Center, New York, New York, USA.

Bioessays : News and Reviews in Molecular, Cellular and Developmental Biology
|July 2, 2022
PubMed

Insights

DNA replication stress disrupts DNA synthesis. A key protein, Rad53, stalls DNA synthesis on both leading and lagging strands, preventing harmful single-stranded DNA formation and maintaining genome integrity.

Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • DNA replication stress poses a significant threat to genome integrity.
  • The conserved DNA replication stress response pathway involves Mec1/ATR, Mrc1/Claspin, and Rad53/Chk1.
  • Leading and lagging DNA strands have distinct protein compositions and functions.

Purpose of the Study:

  • To investigate the strand-specific roles of proteins during DNA replication stress in budding yeast.
  • To analyze protein localization and DNA synthesis at individual DNA strands under replication stress.

Main Methods:

  • Utilized strand-specific sequencing technologies: eSPAN and BrdU-IP-ssSeq.
  • Applied these methods to budding yeast to analyze DNA replication dynamics.

Main Results:

  • Rad53 (an effector kinase) stalls DNA synthesis on both leading and lagging strands during replication stress.
  • Rad53 promotes PCNA unloading on lagging strands.
  • Rad53 attenuates the replication function of Mrc1-Tof1 on leading strands.

Conclusions:

  • Rad53 couples leading and lagging strand DNA synthesis under replication stress.
  • This coupling mechanism prevents the accumulation of potentially harmful single-stranded DNA (ssDNA).
  • The findings elucidate a critical mechanism for maintaining genome stability during replication stress.
Keywords:
ATRDNA replicationDNA replication checkpointMec1Rad53eSPANstrand-specific sequencing

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