title: Threshold effect of growth rate on population variability ofEscherichia colicell lengths.
abstract: A long-standing question in biology is the effect of growth on cell size. Here, we estimate the effect ofEscherichia coligrowth rate (r) on population cell size distributions by estimating the coefficient of variation of cell lengths (CVL) from image analysis of fixed cells in DIC microscopy. We find that the CVLis constant at growth rates less than one division per hour, whereas above this threshold, CVLincreases with an increase in the growth rate. We hypothesize that stochastic inhibition of cell division owing to replication stalling by a RecA-dependent mechanism, combined with the growth rate threshold of multi-fork replication (according to Cooper and Helmstetter), could form the basis of such a threshold effect. We proceed to test our hypothesis by increasing the frequency of stochastic stalling of replication forks with hydroxyurea (HU) treatment and find that cell length variability increases only when the growth rate exceeds this threshold. The population effect is also reproduced in single-cell studies using agar-pad cultures and 'mother machine'-based experiments to achieve synchrony. To test the role of RecA, critical for the repair of stalled replication forks, we examine the CVLofE. coli ΔrecAcells. We find cell length variability in the mutant to be greater than wild-type, a phenotype that is rescued by plasmid-based RecA expression. Additionally, we find that RecA-GFP protein recruitment to nucleoids is more frequent at growth rates exceeding the growth rate threshold and is further enhanced on HU treatment. Thus, we find growth rates greater than a threshold result in increasedE. colicell lengths in the population, and this effect is, at least in part, mediated by RecA recruitment to the nucleoid and stochastic inhibition of division.
authors.affiliation: Division of Biology , Indian Institute of Science Education and Research (IISER) Pune , Dr Homi Bhabha Road, Pashan, Pune 411008 , India.
authors.forename: Manasi S
authors.identifier: 0000-0003-2980-7329
authors.initials: MS
authors.lastname: Gangan
authors.affiliation: Division of Biology , Indian Institute of Science Education and Research (IISER) Pune , Dr Homi Bhabha Road, Pashan, Pune 411008 , India.
authors.forename: Chaitanya A
authors.identifier: 0000-0002-9506-2153
authors.initials: CA
authors.lastname: Athale
keywords: RecA; bacteria; bacterial cell division; cell size; multi-fork replication; population variability