Identification of structural and regulatory cell-shape determinants in Haloferax volcanii

Heather Schiller, Yirui Hong, Joshua Kouassi, Theopi Rados, Jasmin Kwak, Anthony DiLucido, Daniel Safer, Anita Marchfelder, Friedhelm Pfeiffer, Alexandre Bisson (), Stefan Schulze () and Mechthild Pohlschroder ()
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Heather Schiller: University of Pennsylvania, Department of Biology
Yirui Hong: University of Pennsylvania, Department of Biology
Joshua Kouassi: University of Pennsylvania, Department of Biology
Theopi Rados: Brandeis University, Department of Biology
Jasmin Kwak: Brandeis University, Department of Biology
Anthony DiLucido: University of Pennsylvania, Department of Biology
Daniel Safer: University of Pennsylvania, Department of Physiology
Anita Marchfelder: Biology II, Ulm University
Friedhelm Pfeiffer: Biology II, Ulm University
Alexandre Bisson: Brandeis University, Department of Biology
Stefan Schulze: University of Pennsylvania, Department of Biology
Mechthild Pohlschroder: University of Pennsylvania, Department of Biology

Nature Communications, 2024, vol. 15, issue 1, 1-15

Abstract: Abstract Archaea play indispensable roles in global biogeochemical cycles, yet many crucial cellular processes, including cell-shape determination, are poorly understood. Haloferax volcanii, a model haloarchaeon, forms rods and disks, depending on growth conditions. Here, we used a combination of iterative proteomics, genetics, and live-cell imaging to identify mutants that only form rods or disks. We compared the proteomes of the mutants with wild-type cells across growth phases, thereby distinguishing between protein abundance changes specific to cell shape and those related to growth phases. The results identified a diverse set of proteins, including predicted transporters, transducers, signaling components, and transcriptional regulators, as important for cell-shape determination. Through phenotypic characterization of deletion strains, we established that rod-determining factor A (RdfA) and disk-determining factor A (DdfA) are required for the formation of rods and disks, respectively. We also identified structural proteins, including an actin homolog that plays a role in disk-shape morphogenesis, which we named volactin. Using live-cell imaging, we determined volactin’s cellular localization and showed its dynamic polymerization and depolymerization. Our results provide insights into archaeal cell-shape determination, with possible implications for understanding the evolution of cell morphology regulation across domains.

Date: 2024
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DOI: 10.1038/s41467-024-45196-0

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