PMID: 9193003May 30, 1997Paper

Initiation of bacteriophage phi29 DNA replication in vivo: assembly of a membrane-associated multiprotein complex

Journal of Molecular Biology
A Bravo, M Salas

Abstract

Initiation of in vitro phage phi29 DNA replication requires the formation of a heterodimer between a free molecule of terminal protein (TP), which acts as primer, and the viral DNA polymerase. We have analyzed membrane vesicles from phi29-infected Bacillus subtilis cells by quantitative immunoblot techniques. During phage DNA synthesis, large amounts of the viral proteins p1 and free TP were recovered in membrane fractions, as well as a low percentage of the total viral DNA polymerase. Interestingly, the amount of DNA polymerase in membrane fractions increased when viral DNA replication was blocked. Both protein p1 and free TP showed affinity for membranes in the absence of viral DNA. The association of protein p1 with membranes was abolished when the C-terminal 43 amino acid residues were deleted. The above results, together with the critical role of protein p1 for in vivo phi29 DNA replication, led us to conclude that a preliminary stage in the initiation of in vivo phi29 DNA replication could be the assembly of a membrane-associated multiprotein complex containing at least protein p1, free TP and DNA polymerase. Membrane-attachment of this complex could be directly mediated by both protein p1 and free TP. The ability of free...Continue Reading

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Citations

Nov 28, 2000·Journal of Bacteriology·A Camacho, M Salas
Oct 11, 2008·Proceedings of the National Academy of Sciences of the United States of America·Gemma Serrano-HerasMargarita Salas
May 31, 2001·Microbiology and Molecular Biology Reviews : MMBR·W J MeijerM Salas
Aug 9, 2003·The Journal of Biological Chemistry·Gemma Serrano-HerasAlicia Bravo
Jul 10, 2013·Proceedings of the National Academy of Sciences of the United States of America·David Ballesteros-PlazaDaniel Muñoz-Espín
Jun 16, 2015·Frontiers in Microbiology·Julián R DibFriedhelm Meinhardt
Jan 20, 2006·The Journal of Biological Chemistry·Gemma Serrano-HerasAlicia Bravo
Jan 18, 2005·FEMS Microbiology Reviews·Alicia BravoMargarita Salas
Dec 12, 2012·Physical Review Letters·Richard Matthews, Christos N Likos
Dec 23, 2003·Journal of Molecular Biology·Carmen de AntonioManuel Espinosa
Jun 6, 2013·The Journal of Physical Chemistry. B·Richard Matthews, Christos N Likos
Aug 25, 2012·Current Biology : CB·Daniel P Haeusser, William Margolin

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