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Title

Binding of phage phi 29 architectural protein p6 to the viral genome: evidence for topological restriction of the phage linear DNA

AuthorsGonzález-Huici, Víctor; Alcorlo, Martín ; Salas, Margarita ; Hermoso, José Miguel
KeywordsBacillus subtilis
phage phi 29
Issue Date1-Jul-2004
PublisherOxford University Press
CitationNucleic Acids Research 2004 32(11):3493-3502
AbstractBacillus subtilis phage phi 29 protein p6 is required for DNA replication and promotes the switch from early to late transcription. In vivo it binds all along the viral linear DNA, which suggests a global role as an architectural protein; in contrast, binding to bacterial DNA is negligible. This specificity could be due to the p6 binding preference for less negatively supercoiled DNA, as is presumably the case with viral (with respect to bacterial) DNA. Here we demonstrate that p6 binding to phi 29 DNA is greatly increased when negative supercoiling is decreased by novobiocin; in addition, gyrase is required for DNA replication. This indicates that, although non-covalently closed, the viral genome is topologically constrained in vivo. We also show that the p6 binding to different phi 29 DNA regions is modulated by the structural properties of their nucleotide sequences. The higher affinity for DNA ends is possibly related to the presence of sequences in which their bendability properties favor the formation of the p6–DNA complex, whereas the lower affinity for the transcription control region is most probably due to the presence of a rigid intrinsic DNA curvature
Publisher version (URL)http://dx.doi.org/10.1093/nar/gkh668
URIhttp://hdl.handle.net/10261/7841
DOI10.1093/nar/gkh668
ISSN0305-1048
E-ISSN1362-4962
Appears in Collections:(CBM) Artículos
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