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The Bacillus subtilis DNA replication terminator

M T Smith1, C J de Vries, D B Langley

  • 1Department of Biochemistry University of Sydney, NSW, Australia.

Journal of Molecular Biology
|July 5, 1996
PubMed
Summary
This summary is machine-generated.

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Bacillus subtilis replication terminators require two opposing RTP binding sites for function. Manipulating terminator symmetry affects DNA replication fork arrest activity, leading to bidirectional or polar action.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • The discovery of the Bacillus subtilis plasmid terminator TerLS20 highlights the importance of replication terminators.
  • B. subtilis replication terminators, like TerI and TerLS20, feature conserved trinucleotide regions within two 13 nt segments, crucial for DNA replication fork arrest.
  • The interaction of replication terminator protein (RTP) dimers with these sites is key, but not strictly dependent on specific base contacts in the conserved region.

Purpose of the Study:

  • To investigate the structural and functional requirements of B. subtilis replication terminators.
  • To explore the role of symmetry and sequence variations in terminator activity.
  • To model the interaction of RTP dimers with DNA terminators.

Main Methods:

Related Experiment Videos

  • Construction of a synthetic, symmetrical terminator (TerSymB) by modifying the TerI terminator.
  • Assessing the bidirectional and polar fork arrest activity of TerSymB and its variants.
  • Modeling RTP dimer-DNA interactions based on crystal structures and experimental data.

    • Main Results:

    • The synthetic terminator TerSymB confirmed the necessity of two opposing RTP binding sites for functional termination.
    • Sequence deviations in TerSymB converted its activity from bidirectional to predominantly polar.
    • Insertions within TerSymB altered RTP binding and fork arrest, supporting models of dimer positioning and interaction.

    Conclusions:

    • Functional replication terminators in B. subtilis require two symmetrical RTP binding sites.
    • The symmetry and sequence of these sites dictate the directionality of DNA replication fork arrest.
    • A model is proposed to explain the generation of bidirectional or polar fork arrest activity.