Abstract
Gas vesicles consist predominantly of the hydrophobic GvpA and GvpC, and the accessory proteins GvpF through GvpM are required in minor amounts during formation. GvpM and its putative interaction partners were investigated. GvpM interacted with GvpH, GvpJ and GvpL, but not with GvpG. Interactions were also observed in vivo in Haloferax volcanii transformants using Gvp fusions to the green fluorescent protein smGFP. Cells producing the hydrophobic MGFP contained a single fluorescent aggregate per cell, whereas cells containing LGFP or HGFP were fully fluorescent. The soluble LGFP formed stable co-aggregates with GvpM in LGFPM transformants, but the presence of GvpH resulted in the absence of MGFP foci in HMGFP transformants. Substitution- and deletion mutants of GvpM determined functionally important amino acids (aa). Substitution of a polar by a non-polar aa in the N-terminal region of GvpM had no effect, whereas a substitution of a non-polar by a polar aa in this region inhibited gas vesicle formation in transformants. Substitutions in region 44–48 of GvpM strongly reduced the number of gas vesicles, and deletions at the N-terminus resulted in Vac− transformants. Gas vesicle morphology was not affected by any mutation, implying that GvpM is required during initial stages of gas vesicle assembly.
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Chu LJ, Chen MC, Setter J, Tsai YS, Yang H, Fang X, Ting YS, Shaffer GK, Taylor SA, von Haller PD, Goodlett DR, Ng WV (2011) New structural proteins of Halobacterium salinarum gas vesicle revealed by comparative proteomics analysis. J Proteome Res 10:1170–1178
DasSarma S (1993) Identification and analysis of the gas vesicle gene cluster on an unstable plasmid of Halobacterium halobium. Experientia 49:482–486
Englert C, Pfeifer F (1993) Analysis of gas vesicle gene expression in Haloferax mediterranei reveals that GvpA and GvpC are both gas vesicle structural proteins. J Biol Chem 268:9329–9336
Englert C, Krüger K, Offner S, Pfeifer F (1992a) Three different but related gene clusters encoding gas vesicles in halophilic archaea. J Mol Biol 227:586–592
Englert C, Wanner G, Pfeifer F (1992b) Functional analysis of the gas vesicle gene cluster of the halophilic archaeon Haloferax mediterranei defines the vac-region boundary and suggests a regulatory role for the gvpD gene or its product. Mol Microbiol 6:3543–3550
Halladay JT, Jones JG, Lin F, MacDonald AB, DasSarma S (1993) The rightward gas vesicle operon in Halobacterium plasmid pNRC100: identification of the gvpA and gvpC gene products by use of antibody probes and genetic analysis of the region downstream of gvpC. J Bacteriol 175:684–692
Hayes PK, Buchholz B, Walsby AE (1992) Gas vesicles are strengthened by the outer-surface protein, GvpC. Arch Microbiol 157:229–234
Hofacker A, Schmitz KM, Cichonczyk A, Sartorius-Neef S, Pfeifer F (2004) GvpE- and GvpD-mediated transcription regulation of the p-gvp genes encoding gas vesicles in Halobacterium salinarum. Microbiology 150:1829–1838
Lam WL, Doolittle WF (1989) Shuttle vectors for the archaebacterium Halobacterium volcanii. Proc Natl Acad Sci USA 86:5478–5482
Marschaus L, Pfeifer F (2012) A dual promoter region with overlapping activator sequences drives the expression of gas vesicle protein genes in haloarchaea. Microbiology 158:2815–2825
Offner S, Pfeifer F (1995) Complementation studies with the gas vesicle-encoding p-vac region of Halobacterium salinarium PHH1 reveal a regulatory role for the p-gvpDE genes. Mol Microbiol 16:9–19
Offner S, Ziese U, Wanner G, Typke D, Pfeifer F (1998) Structural characteristics of halobacterial gas vesicles. Microbiology 144:1331–1342
Offner S, Hofacker A, Wanner G, Pfeifer F (2000) Eight of fourteen gvp genes are sufficient for formation of gas vesicles in halophilic archaea. J Bacteriol 182:4328–4336
Pfeifer F (2012) Distribution, formation and regulation of gas vesicles. Nat Rev Microbiol 10:705–715
Pfeifer F, Ghahraman P (1993) Plasmid pHH1 of Halobacterium salinarium: characterization of the replicon region, the gas vesicle gene cluster and insertion elements. Mol Gen Genet 238:193–200
Pfeifer F, Offner S, Krüger K, Ghahraman P, Englert C (1994) Transformation of halophilic archaea and investigation of gas vesicle synthesis. Syst Appl Microbiol 16:569–577
Reuter CJ, Maupin-Furlow JA (2004) Analysis of proteasome-dependent proteolysis in Haloferax volcanii cells, using short-lived green fluorescent proteins. Appl Environ Microbiol 70:7530–7538
Sartorius-Neef S, Pfeifer F (2004) In vivo studies on putative Shine-Dalgarno sequences of the halophilic archaeon Halobacterium salinarum. Mol Microbiol 51:579–588
Schägger H, von Jagow G (1987) Tricine-sodium dodecyl sulfate polyacralamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem 166:368–379
Scheuch S, Marschaus L, Sartorius-Neef S, Pfeifer F (2008) Regulation of gvp genes encoding gas vesicle proteins in halophilic Archaea. Arch Microbiol 190:333–339
Schmidt I, Pfeifer F (2013) Use of GFP-GvpE fusions to quantify the GvpD-mediated reduction of the transcriptional activator GvpE in haloarchaea. Arch Microbiol 195:403–412
Shukla HD, DasSarma S (2004) Complexity of gas vesicle biogenesis in Halobacterium sp. strain NRC-1: identification of five new proteins. J Bacteriol 186:3182–3186
Strunk T, Hamacher K, Hoffgaard F, Engelhardt H, Zillig MD, Faist K, Wenzel W, Pfeifer F (2011) Structural model of the gas vesicle protein GvpA and analysis of GvpA mutants in vivo. Mol Microbiol 81:56–68
Tavlaridou S, Faist K, Weitzel K, Pfeifer F (2013) Effect of an overproduction of accessory Gvp proteins on gas vesicle formation in Haloferax volcanii. Extremophiles 17:277–287
Teufel K, Pfeifer F (2010) Interaction of transcription activator GvpE with TATA-box-binding proteins of Halobacterium salinarum. Arch Microbiol 192:143–149
Walsby AE (1994) Gas vesicles. Microbiol Rev 58:94–144
Zimmermann P, Pfeifer F (2003) Regulation of the expression of gas vesicle genes in Haloferax mediterranei: interaction of the two regulatory proteins GvpD and GvpE. Mol Microbiol 49:783–794
Acknowledgements
Karin Faist is thanked for help in electron microscopy. Sabrina Fröls and Arnulf Kletzin are thanked for valuable discussions, and Franz-Josef Meyer-Almes (h_da Darmstadt) for the use of the fluorescence reader. This work received financial support by the German Research Foundation, DFG (Pf 165/12-1).
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Communicated by S. Albers.
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Tavlaridou, S., Winter, K. & Pfeifer, F. The accessory gas vesicle protein GvpM of haloarchaea and its interaction partners during gas vesicle formation. Extremophiles 18, 693–706 (2014). https://doi.org/10.1007/s00792-014-0650-0
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DOI: https://doi.org/10.1007/s00792-014-0650-0