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Requirements for Bacillus subtilis bacteriophage phi29 DNA ejection.

Víctor González-Huici1, Margarita Salas, José M Hermoso

  • 1Instituto de Biología Molecular Eladio Viñuela (CSIC), Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Universidad Autónoma, Cantoblanco, 28049 Madrid, Spain.

Gene
|March 7, 2006
PubMed
Summary

Phage phi29 DNA ejection occurs in two steps. The second step, involving viral protein p17, requires energy and negatively supercoiled DNA, but not transcription.

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Area of Science:

  • Molecular Biology
  • Virology
  • Biochemistry

Background:

  • Phage phi29 infects Bacillus subtilis, ejecting its DNA via a two-step mechanism.
  • The second step of DNA ejection is proposed to involve a motor complex including viral protein p17.

Purpose of the Study:

  • To investigate the energy requirements and molecular mechanisms of phi29 DNA ejection.
  • To elucidate the role of viral protein p17 in the DNA pulling process.

Main Methods:

  • Utilized energy poisons (gramicidin, CCCP, azide) to assess energy dependence.
  • Employed DNA replication and transcription inhibitors (novobiocin) to study regulatory pathways.
  • Analyzed the effect of these agents on phi29 DNA ejection.

Main Results:

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  • The first DNA ejection step is passive and energy-independent.
  • The second step requires an electrochemical proton gradient and is abolished by novobiocin, indicating a need for negatively supercoiled DNA.
  • Viral protein p17 is essential for energy transduction during DNA pulling, as shown by azide's lack of effect in its absence.

Conclusions:

  • Phi29 DNA ejection is a multi-step process with distinct energy requirements for each phase.
  • Viral protein p17 plays a crucial role in harnessing cellular energy for DNA translocation.
  • The mechanism involves DNA supercoiling and proton motive force, highlighting the intricate interplay between phage and host cell machinery.