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Osmotic pressure: resisting or promoting DNA ejection from phage?

Jeembaeva, Meerim LU ; Castelnovo, Martin; Larsson, Frida and Evilevitch, Alex LU (2008) In Journal of Molecular Biology 381(2). p.310-323
Abstract
Recent in vitro experiments have shown that DNA ejection from bacteriophage can be partially stopped by surrounding osmotic pressure when ejected DNA is digested by DNase I in the course of ejection. In this work, we argue by a combination of experimental techniques (osmotic suppression without DNase I monitored by UV absorbance, pulse-field electrophoresis, and cryo-transmission electron microscopy visualization) and simple scaling modeling that intact genome (i.e., undigested) ejection in a crowded environment is, on the contrary, enhanced or eventually complete with the help of a pulling force resulting from DNA condensation induced by the osmotic stress itself. This demonstrates that in vivo, the osmotically stressed cell cytoplasm... (More)
Recent in vitro experiments have shown that DNA ejection from bacteriophage can be partially stopped by surrounding osmotic pressure when ejected DNA is digested by DNase I in the course of ejection. In this work, we argue by a combination of experimental techniques (osmotic suppression without DNase I monitored by UV absorbance, pulse-field electrophoresis, and cryo-transmission electron microscopy visualization) and simple scaling modeling that intact genome (i.e., undigested) ejection in a crowded environment is, on the contrary, enhanced or eventually complete with the help of a pulling force resulting from DNA condensation induced by the osmotic stress itself. This demonstrates that in vivo, the osmotically stressed cell cytoplasm will promote phage DNA ejection rather than resist it. The further addition of DNA-binding proteins under crowding conditions is shown to enhance the extent of ejection. We also found some optimal crowding conditions for which DNA content remaining in the capsid upon ejection is maximum, which correlates well with the optimal conditions of maximum DNA packaging efficiency into viral capsids observed almost 20 years ago. Biological consequences of this finding are discussed. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
osmotic suppression, phage ejection, DNA-binding proteins, PEG, dextran
in
Journal of Molecular Biology
volume
381
issue
2
pages
310 - 323
publisher
Elsevier
external identifiers
  • wos:000258483600007
  • pmid:18602115
  • scopus:47049104754
ISSN
1089-8638
DOI
10.1016/j.jmb.2008.05.081
language
English
LU publication?
yes
id
1bfa11a8-3923-4eb6-8a37-52132b1b474e (old id 1181547)
date added to LUP
2008-10-03 13:02:40
date last changed
2017-01-01 05:40:49
@article{1bfa11a8-3923-4eb6-8a37-52132b1b474e,
  abstract     = {Recent in vitro experiments have shown that DNA ejection from bacteriophage can be partially stopped by surrounding osmotic pressure when ejected DNA is digested by DNase I in the course of ejection. In this work, we argue by a combination of experimental techniques (osmotic suppression without DNase I monitored by UV absorbance, pulse-field electrophoresis, and cryo-transmission electron microscopy visualization) and simple scaling modeling that intact genome (i.e., undigested) ejection in a crowded environment is, on the contrary, enhanced or eventually complete with the help of a pulling force resulting from DNA condensation induced by the osmotic stress itself. This demonstrates that in vivo, the osmotically stressed cell cytoplasm will promote phage DNA ejection rather than resist it. The further addition of DNA-binding proteins under crowding conditions is shown to enhance the extent of ejection. We also found some optimal crowding conditions for which DNA content remaining in the capsid upon ejection is maximum, which correlates well with the optimal conditions of maximum DNA packaging efficiency into viral capsids observed almost 20 years ago. Biological consequences of this finding are discussed.},
  author       = {Jeembaeva, Meerim and Castelnovo, Martin and Larsson, Frida and Evilevitch, Alex},
  issn         = {1089-8638},
  keyword      = {osmotic suppression,phage ejection,DNA-binding proteins,PEG,dextran},
  language     = {eng},
  number       = {2},
  pages        = {310--323},
  publisher    = {Elsevier},
  series       = {Journal of Molecular Biology},
  title        = {Osmotic pressure: resisting or promoting DNA ejection from phage?},
  url          = {http://dx.doi.org/10.1016/j.jmb.2008.05.081},
  volume       = {381},
  year         = {2008},
}