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Polyphosphate granule biogenesis is temporally and functionally tied to cell cycle exit during starvation in Pseudomonas aeruginosa

Racki, Lisa R. and Tocheva, Elitza I. and Dieterle, Michael G. and Sullivan, Meaghan C. and Jensen, Grant J. and Newman, Dianne K. (2017) Polyphosphate granule biogenesis is temporally and functionally tied to cell cycle exit during starvation in Pseudomonas aeruginosa. Proceedings of the National Academy of Sciences of the United States of America, 114 (12). E2440-E2449. ISSN 0027-8424. PMCID PMC5373386. http://resolver.caltech.edu/CaltechAUTHORS:20170306-140446676

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Abstract

Polyphosphate (polyP) granule biogenesis is an ancient and ubiquitous starvation response in bacteria. Although the ability to make polyP is important for survival during quiescence and resistance to diverse environmental stresses, granule genesis is poorly understood. Using quantitative microscopy at high spatial and temporal resolution, we show that granule genesis in Pseudomonas aeruginosa is tightly organized under nitrogen starvation. Following nucleation as many microgranules throughout the nucleoid, polyP granules consolidate and become transiently spatially organized during cell cycle exit. Between 1 and 3 h after nitrogen starvation, a minority of cells have divided, yet the total granule number per cell decreases, total granule volume per cell dramatically increases, and individual granules grow to occupy diameters as large as ∼200 nm. At their peak, mature granules constitute ∼2% of the total cell volume and are evenly spaced along the long cell axis. Following cell cycle exit, granules initially retain a tight spatial organization, yet their size distribution and spacing relax deeper into starvation. Mutant cells lacking polyP elongate during starvation and contain more than one origin. PolyP promotes cell cycle exit by functioning at a step after DNA replication initiation. Together with the universal starvation alarmone (p)ppGpp, polyP has an additive effect on nucleoid dynamics and organization during starvation. Notably, cell cycle exit is temporally coupled to a net increase in polyP granule biomass, suggesting that net synthesis, rather than consumption of the polymer, is important for the mechanism by which polyP promotes completion of cell cycle exit during starvation.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1073/pnas.1615575114DOIArticle
http://www.pnas.org/content/114/12/E2440PublisherArticle
http://www.pnas.org/content/114/12/E2440/suppl/DCSupplementalPublisherSupporting Information
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5373386/PubMed CentralArticle
ORCID:
AuthorORCID
Tocheva, Elitza I.0000-0002-4869-8319
Jensen, Grant J.0000-0003-1556-4864
Newman, Dianne K.0000-0003-1647-1918
Additional Information:© 2017 National Academy of Sciences. Edited by Christine Jacobs-Wagner, Yale University, West Haven, CT, and approved February 6, 2017 (received for review September 21, 2016). Published online before print March 6, 2017. We specially thank Dr. Alasdair McDowall, Howard Hughes Medical Institute, for electron microscopy support. We thank Carol Garland for technical assistance with Energy Dispersive X-ray Spectroscopy (EDS) data collection and analysis using California Institute of Technology (Caltech)’s Applied Physics and Materials Science Department’s Transmission Electron Microscopy Facility. We also thank Brittany J Belin and Noah Ollikainen for help with statistical analysis of the granule position modeling and gratefully acknowledge Megan Bergkessel and other D.K.N. laboratory members for constructive comments on the manuscript. The Caltech Electron Microscopy Facility is funded in part by the Gordon and Betty Moore Foundation, the Agouron Institute, and the Beckman Foundation. This work was funded in part by the Howard Hughes Medical Institute, and the National Institutes of Health (5R01HL117328-03). Lisa Racki is a Damon Runyon Fellow supported by the Damon Runyon Cancer Research Foundation (DRG-2126-12). Author contributions: L.R.R., E.I.T., G.J.J., and D.K.N. designed research; L.R.R., E.I.T., and M.C.S. performed research; L.R.R. contributed new reagents/analytic tools; L.R.R., E.I.T., M.G.D., M.C.S., G.J.J., and D.K.N. analyzed data; and L.R.R. and D.K.N. wrote the paper. The authors declare no conflict of interest. This article is a PNAS Direct Submission. This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1615575114/-/DCSupplemental.
Funders:
Funding AgencyGrant Number
Gordon and Betty Moore FoundationUNSPECIFIED
Agouron InstituteUNSPECIFIED
Arnold and Mabel Beckman FoundationUNSPECIFIED
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
NIH5R01HL117328-03
Damon Runyon Cancer Research FoundationDRG-2126-12
Subject Keywords:polyphosphate; nucleoid; cell cycle; starvation; biomineralization
PubMed Central ID:PMC5373386
Record Number:CaltechAUTHORS:20170306-140446676
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20170306-140446676
Official Citation:Lisa R. Racki, Elitza I. Tocheva, Michael G. Dieterle, Meaghan C. Sullivan, Grant J. Jensen, and Dianne K. Newman Polyphosphate granule biogenesis is temporally and functionally tied to cell cycle exit during starvation in Pseudomonas aeruginosa PNAS 2017 114 (12) E2440-E2449; published ahead of print March 6, 2017, doi:10.1073/pnas.1615575114
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:74801
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:06 Mar 2017 22:24
Last Modified:24 Oct 2017 16:47

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