Abstract

Septins are important components of the cytoskeleton that are highly conserved in eukaryotes and play major roles in cytokinesis, patterning, and many developmental processes. Septins form heteropolymers which assemble into higher-order structures including rings, filaments, and gauzes. In contrast to actin filaments and microtubules, the molecular mechanism by which septins assemble is not well-understood. Here, we report that in the filamentous fungus Aspergillus nidulans, four core septins form heteropolymeric complexes. AspE, a fifth septin lacking in unicellular yeasts, interacts with only one of the core septins, and only during multicellular growth. AspE is required for proper localization of three of the core septins, and requires this same subset of core septins for its own unique cortical localization. The ΔaspE mutant lacks developmentally-specific septin higher-order structures and shows reduced spore production and slow growth with low temperatures and osmotic stress. Our results show that at least two distinct septin heteropolymer populations co-exist in A. nidulans, and that while AspE is not a subunit of either heteropolymer, it is required for assembly of septin higher-order structures found in multicellular development.

Highlights

  • Septin GTPases are key components of the cytoskeleton with roles as important and diverse as those of actin, microtubules and intermediate filaments [1,2,3]

  • Using septin fusion proteins expressed from native septin promoters at wild-type levels, immunoprecipitation, and mass spectrometry, we show that the four A. nidulans core septins interact as expected during early development, but form distinct stage-specific heteropolymers during multicellular growth

  • Our results show that at least two distinct septin heteropolymer classes co-exist in A. nidulans vegetative growth and that while AspE is not required for the formation of either type of heteropolymer, it is required for the association of one class of heteropolymers into higher-order structures specific to multicellular growth

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Summary

Introduction

Septin GTPases are key components of the cytoskeleton with roles as important and diverse as those of actin, microtubules and intermediate filaments [1,2,3]. Septins form heteropolymers, which associate into higher-order structures, typically at the cell cortex, where they can act as diffusion barriers, maintaining proteins within discrete subcellular domains, and scaffolds for reorientation of the F-actin cytoskeleton. The S. cerevisiae septin proteins Cdc, Cdc, Cdc and Cdc were later shown to localize to the yeast neck where they form the 10 nm filament array between the mother cell and daughter bud and serve to restrict and organize cell division proteins at the bud site [4,5,7]

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