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Related Experiment Videos

Replication terminator protein-based replication fork-arrest systems in various Bacillus species

A A Griffiths1, P A Andersen, R G Wake

  • 1Department of Biochemistry, University of Sydney, New South Wales, Australia.

Journal of Bacteriology
|June 27, 1998
PubMed
Summary

Replication terminator protein (RTP) homologs were found in four Bacillus species, revealing conserved function despite genetic variation. This highlights RTP

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • DNA replication requires precise termination to prevent genomic instability.
  • Replication terminator protein (RTP) in Bacillus subtilis arrests replication forks at DNA terminators.
  • Fork-arrest systems in bacteria like E. coli and B. subtilis appear to have evolved independently.

Purpose of the Study:

  • To identify and characterize RTP homologs in species closely related to Bacillus subtilis.
  • To investigate the evolutionary relationships and functional conservation of RTP.
  • To refine the understanding of DNA terminators and the RTP binding mechanism.

Main Methods:

  • Bioinformatic analysis to identify RTP homologs in bacterial genomes.
  • Sequence analysis of nucleotide and amino acid changes.

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  • Phylogenetic analysis to establish evolutionary relationships.
  • Functional assessment of RTP variants in vivo.

    • Main Results:

    • RTP homologs were identified in B. atrophaeus, B. amyloliquefaciens, B. mojavensis, and B. vallismortis, but not B. licheniformis.
    • Conserved amino acid sequences, particularly in the DNA-binding region, suggest functional importance.
    • A naturally occurring variant (E30K) in B. mojavensis RTP functions normally in fork arrest.
    • Identification of new DNA terminators, leading to a refined Bacillus terminator consensus sequence.

    Conclusions:

    • The core function of RTP in replication fork arrest is conserved across closely related Bacillus species.
    • Minimal amino acid changes despite significant nucleotide variation underscore the structural constraints on RTP function.
    • The natural occurrence of the E30K variant challenges previous implications of E30's crucial role in fork arrest.
    • Expanded identification of DNA terminators enhances our understanding of replication termination mechanisms in Bacillus.