Biology subjects

Colarusso, A. V.

Publications and source records attributed to Colarusso, A. V..

4 recordsLinked to original sources

bioRxiv2026

Septins and cytokinesis in the polymorphic fungus Aureobasidium pullulans

During cytokinesis of animals and fungi, a contractile actomyosin ring (CAR) assembles at target locations and constricts to drive cell separation. In animal cells, the position of the CAR is determined by the mitotic spindle, so that the cleavage plane is perpendicular to the mitotic axis. However, in budding yeasts, the location of CAR assembly is specified by a cortical septin cytoskeleton that recruits CAR components to the neck. In the polymorphic fungus Aureobasidium pullulans, we show that septins assemble at mother-bud necks and predict the site of CAR assembly. Cells lacking septins stochastically failed to assemble CARs at a subset of bud necks. However, even cells lacking all four core septins were able to assemble CARs at 75% of bud necks. Our findings suggest the existence of a novel CAR positioning strategy that requires neither septin scaffolds nor nuclear/spindle cues to enable CAR assembly and constriction at bud necks. eTOC SUMMARYBudding yeasts are thought to use septins to mark mother-bud necks as sites for cytokinesis. Here, we find that the multibudding yeast Aureobasidium pullulans can position cytokinetic machinery at most bud necks even in the absence of septins, revealing a novel pathway to mark cytokinesis sites.

cell biology↗

bioRxiv2025

Allocation of resources among multiple daughter cells

Cell division commonly produces two daughter cells, but there are many exceptions where large cells produce multiple daughters. Multiple fission of some green algae and bacteria, cellularization during embryogenesis of plants and insects, and growth of Ichthyosporeans, Chytrids, and Apicomplexans all provide variations on this theme. In some yeast species, a large multi-nucleate mother cell grows multiple buds (daughters) simultaneously. Here we address how mothers partition growth equally among their buds in the multi-budding yeast Aureobasidium pullulans. Bud growth is directed by actin cable networks that appear to be optimized for even partitioning despite complex cell geometries. Even partitioning does not rely on compensatory mechanisms to adjust bud volumes, but rather stems directly from effective equalization of polarity sites. These results reveal how conserved cell polarity and cytoskeletal networks are adapted to build complex morphologies in fungi.

cell biology↗

bioRxiv2025

A genetic strategy to allow detection of F-actin by phalloidin staining in diverse fungi

Actin is highly conserved across eukaryotes. This versatile protein builds cytoskeletal networks central to diverse cellular processes, including cell division and cell motility. The most potent and broadly used reagents to detect polymerized actin distribution in fixed cells are fluorescently conjugated derivatives of the basidiomycete-derived toxin, phalloidin. However, despite its conservation, actin in many ascomycete fungi fails to bind phalloidin. Here we trace the failure to bind phalloidin to a single amino acid change in a phalloidin-binding residue in actin. Reverting this change in the fungus Aureobasidium pullulans by introducing the point mutation act1V75I at the native ACT1 locus confers phalloidin binding without disrupting actin function. We describe a simple genetic technique to introduce this point mutation that may be effective in other fungal systems. This strategy should enable characterization of F-actin in a wider range of fungi. SUMMARYA single point mutation, act1V75I, enables phalloidin staining in ascomycete fungi

cell biology↗

bioRxiv2025

Mechanisms of nuclear segregation in a multinucleate multibudding yeast

Budding yeasts present an especially challenging geometry for segregation of chromosomes, which must be delivered across the narrow mother-bud neck into the bud. Studies in the model yeast Saccharomyces cerevisiae have revealed an elaborate set of mechanisms that selectively orient one mitotic spindle pole towards the bud and then drive spindle elongation along the mother-bud axis, ensuring nuclear segregation between mother and bud. It is unclear how these pathways might be adapted to yield similar precision in more complex cell geometries. Here we provide the first description of the dynamics of mitosis in a multi-nucleate, multibudding yeast, Aureobasidium pullulans, and identify many unexpected differences from uninucleate yeasts. Mitotic spindles do not orient along the mother-bud axis prior to anaphase, and accurate nuclear segregation often occurs after spindle disassembly. Cortical Num1-dynein forces pull highly mobile nuclei into buds, and once a nucleus enters a bud, it discourages others from entering, ensuring that most daughters inherit only one nucleus.

cell biology↗