Protein-primed DNA replication: a transition between two modes of priming by a unique DNA polymerase

The EMBO Journal
J MéndezM Salas

Abstract

Phage phi29 from Bacillus subtilis is a paradigm of the protein-primed replication mechanism, in which a single-subunit DNA polymerase is involved in both the specific protein-primed initiation step and normal DNA elongation. To start phi29 DNA replication, the viral DNA polymerase must interact with a free molecule of the viral terminal protein (TP), to prime DNA synthesis once at each phi29 DNA end. The results shown in this paper demonstrate that the DNA polymerase-primer TP heterodimer is not dissociated immediately after initiation. On the contrary, there is a transition stage in which the DNA polymerase synthesizes a five nucleotide-long DNA molecule while complexed with the primer TP, undergoes some structural change during replication of nucleotides 6-9, and finally dissociates from the primer protein when nucleotide 10 is inserted onto the nascent DNA chain. This behaviour probably reflects the polymerase requirement for a DNA primer of a minimum length to efficiently catalyze DNA elongation. The significance of such a limiting transition stage is supported by the finding of abortive replication products consisting of the primer TP linked up to eight nucleotides, detected during in vitro replication of phi29 TP-DNA par...Continue Reading

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Citations

Nov 9, 2011·Proceedings of the National Academy of Sciences of the United States of America·Mario MencíaMargarita Salas
Apr 25, 2000·Nucleic Acids Research·I GascónM Salas
Jan 20, 2004·Nucleic Acids Research·Verónica TrunigerMargarita Salas
Nov 27, 2008·Nucleic Acids Research·Irene RodríguezMiguel de Vega
May 31, 2001·Microbiology and Molecular Biology Reviews : MMBR·W J MeijerM Salas
Apr 20, 2007·Annual Review of Microbiology·Margarita Salas
Oct 12, 2000·Biochemical and Biophysical Research Communications·K G Ford
Nov 4, 2009·Journal of Molecular Biology·Patricia Pérez-ArnaizMiguel de Vega
Jun 3, 2009·Molecular Microbiology·Tanel Tenson, Vasili Hauryliuk
Aug 11, 2006·The EMBO Journal·Thomas A Steitz
Apr 2, 2008·IUBMB Life·Margarita SalasMiguel de Vega
Apr 6, 2006·FEMS Microbiology Reviews·Christoph Weigel, Harald Seitz
Jan 18, 2005·FEMS Microbiology Reviews·Alicia BravoMargarita Salas
Feb 28, 2015·Nucleic Acids Research·Isabel HolgueraMargarita Salas
Jul 29, 2016·Nucleic Acids Research·Mónica Berjón-OteroModesto Redrejo-Rodríguez
Aug 23, 2016·Frontiers in Molecular Biosciences·Margarita SalasMiguel de Vega
Sep 25, 2015·The Journal of Biological Chemistry·Alicia Del PradoMargarita Salas
May 21, 2019·PloS One·Iván de la TorreAlicia Del Prado
Sep 28, 2000·The Journal of Biological Chemistry·V Gonzalez-HuiciJ M Hermoso
Nov 6, 2012·The Journal of Biological Chemistry·Margarita Salas

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