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Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method
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Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method

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HROB Is Implicated in DNA Replication.

Julia Kutz1,2, Hannes Schmietendorf1,2, Sheikh Anika Rahman1,2

  • 1Project Group Biochemistry, Leibniz Institute on Aging-Fritz Lipmann Institute, D-07745 Jena, Germany.

Genes
|January 8, 2025
PubMed
Summary
This summary is machine-generated.

The newly identified HROB protein interacts with DNA replication machinery and is crucial for maintaining genomic integrity. Its expression peaks during S phase, and its ectopic expression halts DNA replication, suggesting a role in repairing DNA damage.

Keywords:
DNA interstrand crosslink (ICL) repairDNA replicationFörster resonance energy transfer (FRET)HROBprotein overexpressionsoluble protein fragment identification

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • DNA replication is a complex process vital for genetic information duplication.
  • Maintaining genomic integrity requires coordination between DNA damage response, repair, and replication.
  • HROB is a recently identified protein partner and activator of the Mcm8/9 helicase involved in DNA interstrand crosslink (ICL) repair.

Purpose of the Study:

  • To investigate the role of HROB in DNA replication and its association with replication machinery.
  • To understand the expression pattern of HROB during the cell cycle.
  • To elucidate the structural features and evolutionary origin of HROB.

Main Methods:

  • Flow cytometry Förster resonance energy transfer (FRET) assay to detect protein associations.
  • Analysis of HROB expression levels across the cell cycle.
  • Structural prediction and homology searches for HROB.

Main Results:

  • HROB expression is co-regulated with DNA replication factors, peaking in S phase and decreasing in quiescence.
  • HROB is an intrinsically disordered protein with a helix-rich region and an oligonucleotide/oligosaccharide-binding-fold motif.
  • FRET assays confirmed associations between HROB and DNA replication proteins.
  • Ectopic expression of HROB resulted in a near-complete shutdown of DNA replication.

Conclusions:

  • HROB plays a significant role in DNA replication, particularly in navigating challenging DNA structures like ICLs.
  • The findings suggest HROB is a key component in the cellular response to DNA damage during replication.
  • HROB's interaction with the replication machinery highlights its importance in maintaining genomic stability.