LIPID ANCHORING OF ARCHAEOSORTASE SUBSTRATES AND MID-CELL GROWTH IN HALOARCHAEA

Abstract

The archaeal cytoplasmic membrane provides an anchor for many surface proteins. Recently, a novel membrane anchoring mechanism involving a peptidase, archaeosortase A (ArtA) and C-terminal lipid attachment of surface proteins was identified in the model archaeonHaloferax volcanii. ArtA is required for optimal cell growth and morphogenesis, and the S-layer glycoprotein (SLG), the sole component of theH. volcaniicell wall, is one of the targets for this anchoring mechanism. However, how exactly ArtA function and regulation control cell growth and mor-phogenesis is still elusive. Here, we report that archaeal homologs to the bacterial phos-phatidylserine synthase (PssA) and phosphatidylserine decarboxylase (PssD) are involved in ArtA-dependent protein maturation.H. volcaniistrains lacking either HvPssA or HvPssD exhibited motility, growth and morphological phenotypes similar to those of ∆artA. Moreover, we showed the loss of covalent lipid attachment to SLG in the ∆hvpssAmutant and that proteolytic cleavage of the ArtA substrate HVO_0405 was blocked in the ∆hvpssAand ∆hvpssDstrains. Strikingly, ArtA, HvPssA, and HvPssD GFP-fusions co-localized to the mid position ofH. volcaniicells, strongly supporting that they are involved in the same pathway. Finally, we have shown that the SLG is also recruited to the mid cell prior to being secreted and lipid-anchored at the cell outer surface. Collectively, our data suggest haloarchaea use the mid cell as the main surface processing hotspot for cell elongation, division and shape determination.