A conserved molecular machinery centered on the Cdc42 GTPase regulates cell polarity in diverse organisms. Here we review findings from budding and fission yeasts that reveal both a conserved core polarity circuit and several adaptations that each organism exploits to fulfill the needs of its lifestyle. The core circuit involves positive feedback by local activation of Cdc42 to generate a cluster of concentrated GTP-Cdc42 at the membrane. Species-specific pathways regulate the timing of polarization during the cell cycle, as well as the location and number of polarity sites.

/pdf/cell-polarity-in-yeast-4rig25k45p.pdf

Cell Polarity in Yeast.

Mathematical modeling has been instrumental in identifying common principles of cell polarity across diverse systems. These principles include positive feedback loops that are required to destabili...

/pdf/many-roads-to-symmetry-breaking-molecular-mechanisms-and-1o721myzha.pdf

Many roads to symmetry breaking: molecular mechanisms and theoretical models of yeast cell polarity

In yeast and animal cytokinesis, the small guanosine triphosphatase (GTPase) Rho1/RhoA has an established role in formation of the contractile actomyosin ring, but its role, if any, during cleavage-furrow ingression and abscission is poorly understood. Through genetic screens in yeast, we found that either activation of Rho1 or inactivation of another small GTPase, Cdc42, promoted secondary septum (SS) formation, which appeared to be responsible for abscission. Consistent with this hypothesis, a dominant-negative Rho1 inhibited SS formation but not cleavage-furrow ingression or the concomitant actomyosin ring constriction. Moreover, Rho1 is temporarily inactivated during cleavage-furrow ingression; this inactivation requires the protein Cyk3, which binds Rho1-guanosine diphosphate via its catalytically inactive transglutaminase-like domain. Thus, unlike the active transglutaminases that activate RhoA, the multidomain protein Cyk3 appears to inhibit activation of Rho1 (and thus SS formation), while simultaneously promoting cleavage-furrow ingression through primary septum formation. This work suggests a general role for the catalytically inactive transglutaminases of fungi and animals, some of which have previously been implicated in cytokinesis.

/pdf/distinct-roles-of-rho1-cdc42-and-cyk3-in-septum-formation-2flsygd1e7.pdf

Distinct roles of Rho1, Cdc42, and Cyk3 in septum formation and abscission during yeast cytokinesis

Living cells are not always symmetrical. Instead they are often polarized, with a distinct front and back or top and bottom. Cell polarization influences many processes, including how a cell moves and grows, and where it will divide. Breaking symmetry – in other words, making one part of a cell different from the rest – regularly involves a small protein called Cdc42, which can switch between an active and inactive form. This protein is found in a range of organisms from fungi to animals. Budding yeast is a valuable model to study cell polarization. This single-celled fungus polarizes in order to produce a daughter cell or ‘bud’ that emerges out of one end of the mother cell. To become polarized, the mother cell accumulates active Cdc42 in a small area of the cell membrane. This region then becomes the front of the cell, from where the future bud will emerge. However, it is not fully understood how active Cdc42 accumulates at only one place. One model proposed that some molecules of active Cdc42 that are already present on the membrane, recruit polarity proteins that in turn activate other, inactive Cdc42 molecules. This self-amplifying loop could eventually build up a local pool of active Cdc42. However, it has proved challenging to directly test this model. Optogenetics is a technique in which a beam of light is used to manipulate proteins inside cells in a precise manner. The method was first developed in the field of neuroscience over a decade ago, and has more recently been applied to cell biology. Now, Witte et al. have used optogenetics to move polarity proteins to defined places on the membrane of yeast cells and analyse how this change affected the pattern of Cdc42 activation. The experiments showed that polarity proteins not only activate Cdc42 but they also recruit more polarity proteins to the same place. The resulting positive feedback loop leads to active Cdc42 accumulating at one site on the membrane. Further work showed that this mechanism only operates in this manner just before the mother cell replicates its DNA, which is when a yeast cell will normally polarize. These results provide a new perspective on how cells can make one part of the cell different from the rest. Beyond yeast, cell polarization plays a major role when animals, including humans, are developing as embryos or healing wounds. These processes are often controlled by a protein that is equivalent to Cdc42 or by other highly related switch-like proteins. This means that yeast will continue to provide a useful model to study these processes in the laboratory. Lastly, the optogenetics approach developed Witte et al. will be useful to dissect other processes that involve molecules being located at specific place in a cell at specific time.

Cell cycle entry triggers a switch between two modes of Cdc42 activation during yeast polarization

In many cells, morphogenetic events are coordinated with the cell cycle by cyclin-dependent kinases (CDKs). For example, many mammalian cells display extended morphologies during interphase but round up into more spherical shapes during mitosis (high CDK activity) and constrict a furrow during cytokinesis (low CDK activity). In the budding yeast Saccharomyces cerevisiae, bud formation reproducibly initiates near the G1/S transition and requires activation of CDKs at a point called “start” in G1. Previous work suggested that CDKs acted by controlling the ability of cells to polarize Cdc42, a conserved Rho-family GTPase that regulates cell polarity and the actin cytoskeleton in many systems. However, we report that yeast daughter cells can polarize Cdc42 before CDK activation at start. This polarization operates via a positive feedback loop mediated by the Cdc42 effector Ste20. We further identify a major and novel locus of CDK action downstream of Cdc42 polarization, affecting the ability of several other Cdc42 effectors to localize to the polarity site.

https://rupress.org/jcb/article-pdf/218/1/171/1381517/jcb_201806196.pdf

Cell-cycle control of cell polarity in yeast.

The role of Cdc42 and its regulation during cytokinesis is not well understood. Using biochemical and imaging approaches in budding yeast, we demonstrate that Cdc42 activation peaks during the G1/S transition and during anaphase but drops during mitotic exit and cytokinesis. Cdc5/Polo kinase is an important upstream cell cycle regulator that suppresses Cdc42 activity. Failure to down-regulate Cdc42 during mitotic exit impairs the normal localization of key cytokinesis regulators-Iqg1 and Inn1-at the division site, and results in an abnormal septum. The effects of Cdc42 hyperactivation are largely mediated by the Cdc42 effector p21-activated kinase Ste20. Inhibition of Cdc42 and related Rho guanosine triphosphatases may be a general feature of cytokinesis in eukaryotes.

/pdf/inhibition-of-cdc42-during-mitotic-exit-is-required-for-11vkuilnv5.pdf

Benjamin D. Atkins

Papers

Inhibition of Cdc42 during mitotic exit is required for cytokinesis

Symmetry Breaking: Scaffold Plays Matchmaker for Polarity Signaling Proteins