Measurements of DNA lengths remaining in a viral capsid after osmotically suppressed partial ejection

Alex Evilevitch; James W. Gober; Martin Phillips; Charles M. Knobler; William M. Gelbart

doi:10.1529/biophysj.104.045088

Measurements of DNA lengths remaining in a viral capsid after osmotically suppressed partial ejection

Alex Evilevitch, James W. Gober, Martin Phillips, Charles M. Knobler, William M. Gelbart

Research output: Contribution to journal › Article › peer-review

Abstract

The effect of external osmotic pressure on the extent of DNA ejection from bacteriophage-λ was recently investigated (Evilevitch et al., 2003). The total length of DNA ejected was measured via the 260-nm absorption by free nucleotides, after opening of the capsids in the presence of varying amounts of polyethylene glycol 8000 and DNase I. As a function of osmolyte concentration, this absorption was shown to decrease progressively, ultimately vanishing completely for a sufficiently high external osmotic pressure. In this work we report the results of both sedimentation and gel analysis of the length of DNA remaining inside the capsids, as a function of osmolyte concentration. It is confirmed in this way that the progressive inhibition of DNA ejection corresponds to partial ejection from all of the capsids.

Original language	English (US)
Pages (from-to)	751-756
Number of pages	6
Journal	Biophysical journal
Volume	88
Issue number	1
DOIs	https://doi.org/10.1529/biophysj.104.045088
State	Published - Jan 2005
Externally published	Yes

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Biophysics

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10.1529/biophysj.104.045088

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title = "Measurements of DNA lengths remaining in a viral capsid after osmotically suppressed partial ejection",

abstract = "The effect of external osmotic pressure on the extent of DNA ejection from bacteriophage-λ was recently investigated (Evilevitch et al., 2003). The total length of DNA ejected was measured via the 260-nm absorption by free nucleotides, after opening of the capsids in the presence of varying amounts of polyethylene glycol 8000 and DNase I. As a function of osmolyte concentration, this absorption was shown to decrease progressively, ultimately vanishing completely for a sufficiently high external osmotic pressure. In this work we report the results of both sedimentation and gel analysis of the length of DNA remaining inside the capsids, as a function of osmolyte concentration. It is confirmed in this way that the progressive inhibition of DNA ejection corresponds to partial ejection from all of the capsids.",

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note = "Funding Information: The work has been financially supported by the National Science Foundation, through grant No. CHE00-7931 to C.M.K. and grant No. CHE99-88651 to W.M.G., and by the Swedish Foundation for Internationalization of Research and Higher Education through a grant to A.E.",

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AU - Evilevitch, Alex

AU - Gober, James W.

AU - Phillips, Martin

AU - Knobler, Charles M.

AU - Gelbart, William M.

N1 - Funding Information: The work has been financially supported by the National Science Foundation, through grant No. CHE00-7931 to C.M.K. and grant No. CHE99-88651 to W.M.G., and by the Swedish Foundation for Internationalization of Research and Higher Education through a grant to A.E.

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N2 - The effect of external osmotic pressure on the extent of DNA ejection from bacteriophage-λ was recently investigated (Evilevitch et al., 2003). The total length of DNA ejected was measured via the 260-nm absorption by free nucleotides, after opening of the capsids in the presence of varying amounts of polyethylene glycol 8000 and DNase I. As a function of osmolyte concentration, this absorption was shown to decrease progressively, ultimately vanishing completely for a sufficiently high external osmotic pressure. In this work we report the results of both sedimentation and gel analysis of the length of DNA remaining inside the capsids, as a function of osmolyte concentration. It is confirmed in this way that the progressive inhibition of DNA ejection corresponds to partial ejection from all of the capsids.

AB - The effect of external osmotic pressure on the extent of DNA ejection from bacteriophage-λ was recently investigated (Evilevitch et al., 2003). The total length of DNA ejected was measured via the 260-nm absorption by free nucleotides, after opening of the capsids in the presence of varying amounts of polyethylene glycol 8000 and DNase I. As a function of osmolyte concentration, this absorption was shown to decrease progressively, ultimately vanishing completely for a sufficiently high external osmotic pressure. In this work we report the results of both sedimentation and gel analysis of the length of DNA remaining inside the capsids, as a function of osmolyte concentration. It is confirmed in this way that the progressive inhibition of DNA ejection corresponds to partial ejection from all of the capsids.

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Measurements of DNA lengths remaining in a viral capsid after osmotically suppressed partial ejection

Abstract

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