Abstract
Coordination of growth and division in eukaryotic cells is essential for populations of proliferating cells to maintain size homeostasis, but the underlying mechanisms that govern cell size have only been investigated in a few taxa. The green alga Chlamydomonas reinhardtii (Chlamydomonas) proliferates using a multiple fission cell cycle that involves a long G1 phase followed by a rapid series of successive S and M phases (S/M) that produces 2n daughter cells. Two control points show cell-size dependence: Commitment in mid-G1 phase requires attainment of a minimum size to enable at least one mitotic division during S/M, and the S/M control point where mother cell size governs cell division number (n), ensuring that daughter distributions are uniform. tny1 mutants pass Commitment at a smaller size than wild type and undergo extra divisions during S/M phase to produce small daughters, indicating that TNY1 functions to inhibit size-dependent cell cycle progression. TNY1 encodes a cytosolic hnRNP A- related RNA binding protein and is produced once per cell cycle during S/M phase where it is apportioned to daughter cells, and then remains at constant absolute abundance as cells grow, a property known as subscaling (1). Altering the dosage of TNY1 in heterozygous diploids or through overexpression increased Commitment cell size and daughter cell size, indicating that TNY1 is a limiting factor for both size control checkpoints. Epistasis placed TNY1 function upstream of the retinoblastoma tumor suppressor complex (RBC) and one of its regulators, Cyclin-Dependent Kinase G1 (CDKG1) (2). Moreover, CDKG1 protein and mRNA were found to over-accumulate in tny1 cells suggesting that CDKG1 may be a direct target of repression by TNY1. Our data expand the potential roles of subscaling proteins outside the nucleus and imply a control mechanism that ties TNY1 accumulation to pre-division mother cell size.
Author Summary Size control is a fundamental property of cells which requires balancing cell growth with cell division, but the mechanisms used by cells to achieve this balance are only partly understood. The best-characterized mechanisms for size control to date involve fixed amounts of nuclear- DNA-bound inhibitory factors which repress cell division until cells grow past a minimum size threshold to overcome the inhibition. The unicellular green alga Chlamydomonas and many other algae and protists use a non-canonical cell cycle where cells can grow by many-fold in size before dividing, and then undergo multiple fission which involves successive rapid divisions to produce a uniform-sized population of daughters. In Chlamydomonas an unknown size homeostasis mechanism couples mother cell size to division number such that larger mother cells divide more times than smaller mother cells. Here, we identified and characterized a key factor governing size control in Chlamydomonas, a cytoplasmic RNA-binding protein and division inhibitor, TNY1, that is produced in a fixed amount in daughter cells and does not increase with cell growth, a property called sub-scaling. We found that TNY1 represses production of a cell cycle activator, CDKG1, during multiple fission to control daughter cell size. TNY1 is the first example of a cytosolic cell cycle inhibitor that does not depend on nuclear DNA binding to govern sub-scaling.
Competing Interest Statement
The authors have declared no competing interest.
