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Journal of Bacteriology, December 2004, p. 8401-8406, Vol. 186, No. 24
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.24.8401-8406.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Phage {phi}29 Proteins p1 and p17 Are Required for Efficient Binding of Architectural Protein p6 to Viral DNA In Vivo

Víctor González-Huici, Martín Alcorlo, Margarita Salas,* and José M. Hermoso

Instituto de Biología Molecular "Eladio Viñuela" (CSIC), Centro de Biología Molecular "Severo Ochoa" (CSIC-UAM), Universidad Autónoma, Canto Blanco, Madrid, Spain

Received 12 April 2004/ Accepted 1 September 2004

Bacteriophage {phi}29 protein p6 is a viral architectural protein, which binds along the whole linear {phi}29 DNA in vivo and is involved in initiation of DNA replication and transcription control. Protein p1 is a membrane-associated viral protein, proposed to attach the viral genome to the cell membrane. Protein p17 is involved in pulling {phi}29 DNA into the cell during the injection process. We have used chromatin immunoprecipitation and real-time PCR to analyze in vivo p6 binding to DNA in cells infected with {phi}29 sus1 or sus17 mutants; in both cases p6 binding is significantly decreased all along {phi}29 DNA. {phi}29 DNA is topologically constrained in vivo, and p6 binding is highly increased in the presence of novobiocin, a gyrase inhibitor that produces a loss of DNA negative superhelicity. Here we show that, in cells infected with {phi}29 sus1 or sus17 mutants, the increase of p6 binding by novobiocin is even higher than in cells containing p1 and p17, alleviating the p6 binding deficiency. Therefore, proteins p1 and p17 could be required to restrain the proper topology of {phi}29 DNA, which would explain the impaired DNA replication observed in cells infected with sus1 or sus17 mutants.

* Corresponding author. Mailing address: Instituto de Biología Molecular "Eladio Viñuela" (CSIC), Centro de Biología Molecular "Severo Ochoa" (CSIC-UAM), Universidad Autónoma, Canto Blanco, 28049 Madrid, Spain. Phone: 34-914978435. Fax: 34-914978490. E-mail: msalas{at}cbm.uam.es.

Journal of Bacteriology, December 2004, p. 8401-8406, Vol. 186, No. 24
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.24.8401-8406.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Alcorlo, M., Gonzalez-Huici, V., Hermoso, J. M., Meijer, W. J. J., Salas, M. (2007). The Phage {phi}29 Membrane Protein p16.7, Involved in DNA Replication, Is Required for Efficient Ejection of the Viral Genome. J. Bacteriol. 189: 5542-5549 [Abstract] [Full Text]  


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