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Challenging Packaging Limits and Infectivity of Phage λ

Nurmemmedov, Elmar LU ; Castelnovo, Martin; Medina, Elizabeth; Catalano, Carlos Enrique and Evilevitch, Alex LU (2012) In Journal of Molecular Biology 415(2). p.263-273
Abstract
The terminase motors of bacteriophages have been shown to be among the strongest active machines in the biomolecular world, being able to package several tens of kilobase pairs of viral genome into a capsid within minutes. Yet, these motors are hindered at the end of the packaging process by the progressive buildup of a force-resisting packaging associated with already packaged DNA. In this experimental work, we raise the issue of what sets the upper limit on the length of the genome that can be packaged by the terminase motor of phage λ and still yield infectious virions and the conditions under which this can be efficiently performed. Using a packaging strategy developed in our laboratory of building phage λ from scratch, together with... (More)
The terminase motors of bacteriophages have been shown to be among the strongest active machines in the biomolecular world, being able to package several tens of kilobase pairs of viral genome into a capsid within minutes. Yet, these motors are hindered at the end of the packaging process by the progressive buildup of a force-resisting packaging associated with already packaged DNA. In this experimental work, we raise the issue of what sets the upper limit on the length of the genome that can be packaged by the terminase motor of phage λ and still yield infectious virions and the conditions under which this can be efficiently performed. Using a packaging strategy developed in our laboratory of building phage λ from scratch, together with plaque assay monitoring, we have been able to show that the terminase motor of phage λ is able to produce infectious particles with up to 110% of the wild-type λ-DNA length. However, the phage production rate, and thus the infectivity, decreased exponentially with increasing DNA length and was a factor of 10(3) lower for the 110% λ-DNA phage. Interestingly, our in vitro strategy was still efficient in fully packaging phages with DNA lengths as high as 114% of the wild-type length, but these viruses were unable to infect bacterial cells efficiently. Further, we demonstrated that the phage production rate is modulated by the presence of multivalent ionic species. The biological consequences of these findings are discussed. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
pressure, terminase, DNA packaging, bacteriophage lambda
in
Journal of Molecular Biology
volume
415
issue
2
pages
263 - 273
publisher
Elsevier
external identifiers
  • wos:000300032500003
  • pmid:22108169
  • scopus:84855819055
ISSN
1089-8638
DOI
10.1016/j.jmb.2011.11.015
language
English
LU publication?
yes
id
845369a6-2d06-4a67-93de-de652215979b (old id 2220527)
date added to LUP
2011-12-13 16:46:43
date last changed
2017-01-08 04:24:03
@article{845369a6-2d06-4a67-93de-de652215979b,
  abstract     = {The terminase motors of bacteriophages have been shown to be among the strongest active machines in the biomolecular world, being able to package several tens of kilobase pairs of viral genome into a capsid within minutes. Yet, these motors are hindered at the end of the packaging process by the progressive buildup of a force-resisting packaging associated with already packaged DNA. In this experimental work, we raise the issue of what sets the upper limit on the length of the genome that can be packaged by the terminase motor of phage λ and still yield infectious virions and the conditions under which this can be efficiently performed. Using a packaging strategy developed in our laboratory of building phage λ from scratch, together with plaque assay monitoring, we have been able to show that the terminase motor of phage λ is able to produce infectious particles with up to 110% of the wild-type λ-DNA length. However, the phage production rate, and thus the infectivity, decreased exponentially with increasing DNA length and was a factor of 10(3) lower for the 110% λ-DNA phage. Interestingly, our in vitro strategy was still efficient in fully packaging phages with DNA lengths as high as 114% of the wild-type length, but these viruses were unable to infect bacterial cells efficiently. Further, we demonstrated that the phage production rate is modulated by the presence of multivalent ionic species. The biological consequences of these findings are discussed.},
  author       = {Nurmemmedov, Elmar and Castelnovo, Martin and Medina, Elizabeth and Catalano, Carlos Enrique and Evilevitch, Alex},
  issn         = {1089-8638},
  keyword      = {pressure,terminase,DNA packaging,bacteriophage lambda},
  language     = {eng},
  number       = {2},
  pages        = {263--273},
  publisher    = {Elsevier},
  series       = {Journal of Molecular Biology},
  title        = {Challenging Packaging Limits and Infectivity of Phage λ},
  url          = {http://dx.doi.org/10.1016/j.jmb.2011.11.015},
  volume       = {415},
  year         = {2012},
}