Dilution of the cell cycle inhibitor Whi5 controls budding yeast cell size

Cell size fundamentally affects all biosynthetic processes by determining the scale of organelles and influencing surface transport(1,2). Although extensive studies have identified many mutations affecting cell size, the molecular mechanisms underlying size control have remained elusive(3). In budding yeast, size control occurs in G1 phase prior to Start, the point of irreversible commitment to cell division(4,5). It was previously thought that activity of the G1 cyclin Cln3 increased with cell size to trigger Start by initiating the inhibition of the transcriptional inhibitor Whi5(6-8). However, while Cln3 concentration does modulate the rate at which cells pass Start, we found that its synthesis increases in proportion to cell size so that its total concentration is nearly constant during pre-Start G1. Rather than increasing Cln3 activity, we identify decreasing Whi5 activity — due to the dilution of Whi5 by cell growth — as a molecular mechanism through which cell size controls proliferation. Whi5 is synthesized in S/G2/M phases of the cell cycle in a largely size-independent manner. This results in smaller daughter cells being born with higher Whi5 concentrations that extend their pre-Start G1 phase. Thus, at its most fundamental level, budding yeast size control results from the differential scaling of Cln3 and Whi5 synthesis rates with cell size. More generally, our work shows that differential size-dependency of protein synthesis can provide an elegant mechanism to coordinate cellular functions with growth.