Self-Consistent Examination of Donachie's Constant Initiation Size at the Single-Cell Level.

How growth, the cell cycle, and cell size are coordinated is a fundamental question in biology. Recently, we and others have shown that bacterial cells grow by a constant added size per generation, irrespective of the birth size, to maintain size homeostasis. This "adder" principle raises a question as to when during the cell cycle size control is imposed. Inspired by this question, we examined our single-cell data for initiation size by employing a self-consistency approach originally used by Donachie. Specifically, we assumed that individual cells divide after constant C + D minutes have elapsed since initiation, independent of the growth rate. By applying this assumption to the cell length vs. time trajectories from individual cells, we were able to extract theoretical probability distribution functions for initiation size for all growth conditions. We found that the probability of replication initiation shows peaks whenever the cell size is a multiple of a constant unit size, consistent with the Donachie's original analysis at the population level. Our self-consistent examination of the single-cell data made experimentally testable predictions, e.g., two consecutive replication cycles can be initiated during a single cell-division cycle.