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A Single-Cell Approach Reveals Variation in Cellular Phage-Producing Capacities

Sherin Kannoly, Gabriella Oken, Jon Shadan, David Musheyev, Kevin Singh, Abhyudai Singh, View ORCID ProfileJohn J. Dennehy
doi: https://doi.org/10.1101/2021.10.19.465070
Sherin Kannoly
1Biology Department, Queens College of The City University of New York, NYC, NY, USA
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Gabriella Oken
1Biology Department, Queens College of The City University of New York, NYC, NY, USA
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Jon Shadan
1Biology Department, Queens College of The City University of New York, NYC, NY, USA
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David Musheyev
1Biology Department, Queens College of The City University of New York, NYC, NY, USA
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Kevin Singh
1Biology Department, Queens College of The City University of New York, NYC, NY, USA
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Abhyudai Singh
2Department of Electrical and Computer Engineering, University of Delaware, Newark, DE, USA
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John J. Dennehy
1Biology Department, Queens College of The City University of New York, NYC, NY, USA
3The Graduate Center of The City University of New York, NYC, NY, USA
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  • ORCID record for John J. Dennehy
  • For correspondence: john.dennehy@qc.cuny.edu
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ABSTRACT

Virus burst size, a component of viral fitness, is the average number of viral particles released from a single infected cell. In this study, we estimated bacteriophage lambda (λ) burst size mean and distribution at different lysis times. To estimate phage λ burst sizes at single-cell level, we employed a lysis-deficient E. coli lysogen, which allowed chemical lysis at desired times after the induction of lytic cycle. Induced cultures of E. coli lysogen were diluted and aliquoted into the wells of 96-well plates. A high dilution rate results in mostly empty wells and minimizes the probability of having multiple cells in wells that do receive cells. Burst size was estimated by titering single-cell lysates obtained after chemical lysis at desired times. Our data shows that the viral burst size initially increases exponentially with the lysis time, and then saturates at longer lysis times. We also demonstrate that cell-to-cell variation or “noise” in lysis timing does not significantly contribute to the burst size noise. The burst size noise remains constant with increasing mean burst size. The most likely explanation for the experimentally observed constant burst size noise is that cell-to-cell differences in burst size originate from differences in cellular capacity to produce phages. The mean burst size measured at different lysis times is positively correlated to cell volume, which may determine the cellular phage production capacity. However, experiments controlling for cell size indicates that there are other factors in addition to cell size that determine this cellular capacity.

ARTICLE IMPORTANCE Phages produce offspring by hijacking a cell’s replicative machinery. Previously, it was noted that the variation in the number of phages produced by single infected cells far exceeded cell size variation. It has been hypothesized that this variation is a consequence of variation in the timing of host cell lysis. Here we show that cell-to-cell variation in lysis timing does not significantly contribute to the burst size variation. We suggest that the constant burst size variation across different host lysis times results from cell-to-cell differences in capacity to produce phages. We find that the mean burst size measured at different lysis times is positively correlated to cell volume, which may determine the cellular phage production capacity. However, experiments controlling for cell size indicates that there are other factors in addition to cell size that determine this cellular capacity.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-ND 4.0 International license.
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Posted October 20, 2021.
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A Single-Cell Approach Reveals Variation in Cellular Phage-Producing Capacities
Sherin Kannoly, Gabriella Oken, Jon Shadan, David Musheyev, Kevin Singh, Abhyudai Singh, John J. Dennehy
bioRxiv 2021.10.19.465070; doi: https://doi.org/10.1101/2021.10.19.465070
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A Single-Cell Approach Reveals Variation in Cellular Phage-Producing Capacities
Sherin Kannoly, Gabriella Oken, Jon Shadan, David Musheyev, Kevin Singh, Abhyudai Singh, John J. Dennehy
bioRxiv 2021.10.19.465070; doi: https://doi.org/10.1101/2021.10.19.465070

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