CRISPR-mediated gene silencing reveals involvement of the archaeal S-layer in cell division and virus infection

Isabelle Anna Zink, Kevin Pfeifer, Erika Wimmer, Uwe B. Sleytr, Bernhard Schuster and Christa Schleper ()
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Isabelle Anna Zink: University of Vienna
Kevin Pfeifer: University of Vienna
Erika Wimmer: University of Vienna
Uwe B. Sleytr: University of Natural Resources and Life Sciences
Bernhard Schuster: University of Natural Resources and Life Sciences
Christa Schleper: University of Vienna

Nature Communications, 2019, vol. 10, issue 1, 1-14

Abstract: Abstract The S-layer is a proteinaceous surface lattice found in the cell envelope of bacteria and archaea. In most archaea, a glycosylated S-layer constitutes the sole cell wall and there is evidence that it contributes to cell shape maintenance and stress resilience. Here we use a gene-knockdown technology based on an endogenous CRISPR type III complex to gradually silence slaB, which encodes the S-layer membrane anchor in the hyperthermophilic archaeon Sulfolobus solfataricus. Silenced cells exhibit a reduced or peeled-off S-layer lattice, cell shape alterations and decreased surface glycosylation. These cells barely propagate but increase in diameter and DNA content, indicating impaired cell division; their phenotypes can be rescued through genetic complementation. Furthermore, S-layer depleted cells are less susceptible to infection with the virus SSV1. Our study highlights the usefulness of the CRISPR type III system for gene silencing in archaea, and supports that an intact S-layer is important for cell division and virus susceptibility.

Date: 2019
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DOI: 10.1038/s41467-019-12745-x

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