Interaction of two strongly divergent archaellins stabilizes the structure of the Halorubrum archaellum

Pyatibratov, M. G.; Syutkin, A. S.; Quax, T. E. F.; Melnik, T. N.; Papke, R. T.; Gogarten, J. P.; Kireev, I. I.; Surin, A. K.; Beznosov, S. N.; Galeva, A. V.; Fedorov, O. V.

doi:10.1101/836379

bioRxiv · 10.1101/836379

Interaction of two strongly divergent archaellins stabilizes the structure of the Halorubrum archaellum

Pyatibratov, M. G.·Syutkin, A. S.·Quax, T. E. F.·Melnik, T. N.·Papke, R. T.·Gogarten, J. P.·Kireev, I. I.·Surin, A. K.·Beznosov, S. N.·Galeva, A. V.·Fedorov, O. V.

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Abstract

The archaellum is a unique motility structure that has only functional similarity to its bacterial counterpart, the flagellum. Archaellar filaments consist of thousands of copies of the protein protomer archaellin. Most euryarchaeal genomes encode multiple homologous archaellins. The role of these multiple archaellin genes remains unclear. Halophilic archaea from the genus Halorubrum possess two archaellin genes, flaB1 and flaB2. Amino acid sequences of the corresponding protein products are extraordinarily diverged (identity of [~] 40%). To clarify roles for each archaellin, we compared archaella from two natural Halorubrum lacusprofundi strains: the DL18 strain, which possesses both archaellin genes, and the type strain ACAM 34 whose genome contains the flaB2 gene only. Both strains synthesize functional archaella; however, the DL18 strain, where both archaellins are present in comparable amounts, is more motile. In addition, we expressed these different Hrr. lacusprofundi archaellins in a Haloferax volcanii strain from which the endogenous archaellin genes were deleted. Three Hfx. volcanii strains expressing Hrr. lacusprofundi archaellins flaB1, flaB2 or flaB1-flaB2 produced archaellum filaments consisting of only one (FlaB1 or FlaB2) or both (FlaB1/B2) archaellins. All three recombinant Hfx. volcanii strains were motile, although there were profound differences in the efficiency of motility. The recombinant filaments resemble the natural filaments of Hrr. lacusprofundi. Electron microscopy showed that FlaB1 FlaB2-archaella look like typical supercoiled filaments, while with the shape of the FlaB1- and FlaB2-archaella is more variable. Both native and recombinant FlaB1 FlaB2-filaments have greater thermal stability and are more resistant to low salinity stress than single-component filaments. This shows that thermal stability of archaellins depends on the presence of both archaellin types, indicating a close interaction between these subunits in the supramolecular structure. Functional helical Hrr. lacusprofundi archaella can be composed of either single archaellin: FlaB2 or FlaB1; however, the two divergent archaellin subunits in combination provide additional stabilization to the archaellum structure and thus adaptation to a wider range of external conditions. A comparative genomic analysis of archaellins suggests that the described combination of divergent archaellins is not restricted to Hrr. lacusprofundi, but is occurring also in organisms from other haloarchaeal genera.

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Pyatibratov, M. G., Syutkin, A. S., Quax, T. E. F., Melnik, T. N., Papke, R. T., Gogarten, J. P., Kireev, I. I., Surin, A. K., Beznosov, S. N., Galeva, A. V., Fedorov, O. V.. 2019-11-09. Interaction of two strongly divergent archaellins stabilizes the structure of the Halorubrum archaellum. https://doi.org/10.1101/836379

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