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Annual Review of Virology

Volume 3, 2016

Single-Cell Studies of Phage λ: Hidden Treasures Under Occam's Rug

Abstract

Studies over more than half a century have resulted in what some consider a complete narrative for the life cycle of bacteriophage λ. However, this narrative is only complete within the limited resolution offered by the traditional genetic and biochemical approaches that were used to create it. A recent series of studies performed at the single-cell and single-phage levels has revealed a wealth of previously unknown features. By pointing to many open questions, these new studies highlight the limitations of our current understanding of λ, but they also initiate the process of forming a more detailed and quantitative narrative for the system.

Keyword(s): decision-making, Escherichia coli, fluorescence microscopy, lysis, lysogeny, noise, phage lambda, systems biology

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2016-09-29
2024-04-18
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Single-Cell Studies of Phage λ: Hidden Treasures Under Occam's Rug
Annual Review of Virology 3, 453 (2016); https://doi.org/10.1146/annurev-virology-110615-042127
/content/journals/10.1146/annurev-virology-110615-042127
/content/journals/10.1146/annurev-virology-110615-042127
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Supplemental Material

    Cell lysis following λ infection, as seen under phase-contrast illumination (see also ). The cell is infected at = 0 and lyses at ≈ 80 min. Video courtesy of Louis McLane and Samuel Skinner; Golding lab (unpublished).

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    Tracking the postinfection decision (see also ). Phage capsids were labeled using gpD-YFP. At = 0, two cells are each infected by a single phage (), and one cell is infected by three phages. The two cells infected by single phages go into the lytic pathway, as indicated by the intracellular production of new phage capsids (). The cell infected by three phages goes into the lysogenic pathway, as indicated by the activity of a P-mCherry reporter (). Eventually, the lytic pathway results in cell lysis, whereas the lysogenic cell continues to grow and divide. Video reproduced from Reference 12, copyright 2010, with permission from Elsevier.

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    P activity during lysogen induction (see also ). In the reporter strain, mRNA from the P promoter is labeled using MS2-GFP (), whereas the genome locus where the reporter resides is labeled using TetR-mCherry (). The cells are λ lysogens. At = 20 min, mitomycin C is added, leading to transient derepression of P and induction of the dormant prophage. The prophage used is lysis deficient; hence, the cells do not lyse at the end of the induction process. Video courtesy of Jing Zhang, Louis McLane, and Samuel Skinner; Golding lab (unpublished).

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    Single-molecule imaging of CI production () during stable lysogeny. For more information see Reference 35. Video reproduced from Reference 35, copyright 2012, with permission from Macmillan Publishers Ltd.

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    A single gpD-YFP-labeled λ phage () diffusing near and on the cell (). Two examples are shown sequentially. For more information see Reference 9. Video courtesy of Eli Rothenberg, Samuel Skinner, and Ido Golding (unpublished).

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    DNA injection into the cell. The λ genome, initially inside the capsid, was labeled using a SYTOX Orange DNA dye. For more information see Reference 10. Video reproduced from Reference 10, copyright 2012, with permission from Elsevier.

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    Spatiotemporal dynamics of phage () and bacterial () genome loci inside the infected cell during the lysogenic pathway (see ). The phage genome is labeled using mCherry-ParB. In the same cell, the bacterial site is detected using an orthogonal ParB system and GFP. For more information see Reference 11. Video courtesy of Louis McLane and Samuel Skinner; Golding lab (unpublished).

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    Spatiotemporal dynamics of the phage genome inside the cell during the lytic pathway. At = 0 min, the cell is infected by a gpD-YFP-labeled phage (). At = 5 min, a SeqA-ECFP focus corresponding to the injected phage DNA appears. Later, this focus is converted to two foci, corresponding to two hemimethylated phage DNAs. The cell eventually lyses (not shown). Video reproduced from Reference 38, copyright 2015, with permission from Elsevier.

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  • Article Type: Review Article

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