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The integral membrane protein p16.7 organizes in vivo phi29 DNA replication through interaction with both the

Alejandro Serna-Rico1, Daniel Muñoz-Espín, Laurentino Villar

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

The EMBO Journal
|May 3, 2003
PubMed

Insights

The Bacillus subtilis phage phi29 protein p16.7 forms multimers, crucial for its DNA binding during membrane-associated replication. This study reveals key structural and functional insights into prokaryotic DNA replication organization.

Area of Science:

  • Molecular Biology
  • Virology
  • Cellular Biology

Background:

  • Prokaryotic DNA replication, particularly membrane-associated processes, remains poorly understood.
  • The Bacillus subtilis phage phi29 system offers a model for studying these mechanisms.
  • The phi29-encoded integral membrane protein p16.7 was previously identified as a ssDNA binder involved in replication organization.

Purpose of the Study:

  • To investigate the multimerization properties of the phi29 p16.7 protein.
  • To elucidate the role of p16.7 multimerization in its interaction with DNA and other viral proteins.
  • To develop a model for the in vivo organization of membrane-associated phi29 DNA replication.

Main Methods:

  • In vitro and in vivo multimerization assays of p16.7.
  • Analysis of p16.7 interactions with single-stranded DNA (ssDNA) and the phi29 terminal protein.
  • Site-directed mutagenesis to identify regions involved in multimerization.

Main Results:

  • p16.7 forms multimers both in vitro and in vivo.
  • Multimerization is enhanced by the presence of ssDNA.
  • The C-terminal region of p16.7 is essential for multimerization but not for ssDNA binding or terminal protein interaction.
  • p16.7 multimerization is critical for its ssDNA-binding mode.

Conclusions:

  • p16.7 functions as a multimeric protein in phi29 DNA replication.
  • The protein possesses distinct functional modules, including those for multimerization, ssDNA binding, and terminal protein interaction.
  • A model is proposed integrating p16.7's structure and function in organizing membrane-associated viral DNA replication.

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