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

Archaea are key players in various processes, yet little is known about their cell biology, including cell-shape determination. Haloferax volcanii is a haloarchaeon that can form both rod- and disk-shaped cells, depending on growth conditions. In this study, we used proteomics, genetics, and live-cell imaging to identify mutants that are defective in rod or disk formation. Diverse proteins were implicated to be important for cell-shape determination, including predicted transporters, transducers, signaling molecules, and transcriptional regulators. We identified and phenotypically characterized two deletion strains, strains missing rod-determining factor A (RdfA) and disk-determining factor A (DdfA), which are required for rod and disk formation, respectively. We also identified an actin homolog, volactin, which we characterized with live-cell imaging and determined to be dynamic and important for disk-shape determination.This Zenodo project includes the CellProfiler pipeline (.cpproj file) used for analyzing and quantifying cell-shape images. Additionally, the cell-shape images taken during the proteomics sample collection (comparing H53, ∆rdfA, and ∆ddfA at early- and late-log growth phases) are provided. This project also includes whole genome sequencing Illumina files for H53, ∆rdfA, ∆sph3, ∆ddfA, JK3, and JK5.These data accompany:Schiller, H., Hong, Y., Kouassi, J., Rados, T., Kwak, J., DiLucido, A., Safer, D., Marchfelder, A., Pfeiffer, F., Bisson, A., Schulze, S., and Pohlschroder, M. 2024. Identification of structural and regulatory cell-shape determinants in Haloferax volcanii. Nat Commun 15(1):1414. https://doi.org/10.1038/s41467-024-45196-0.