Complexity of Gas Vesicle Biogenesis in Halobacterium sp. Strain NRC-1: Identification of Five New Proteins

Hem Shukla, Shiladitya DasSarma

Research output: Contribution to journalArticle

Abstract

The genome of Halobacterium sp. strain NRC-1 contains a large gene cluster, gvpMLKJIHGFEDACNO, that is both necessary and sufficient for the production of buoyant gas-filled vesicles. Due to the resistance of gas vesicles to solubilization, only the major gas vesicle protein GvpA and a single minor protein, GvpC, were previously detected. Here, we used immunoblotting analysis to probe for the presence of gas vesicle proteins corresponding to five additional gvp gene products. Polyclonal antisera were raised in rabbits against LacZ-GvpF, -GvpJ, and -GvpM fusion proteins and against synthetic 15-amino-acid peptides from GvpG and -L. Immunoblotting analysis was performed on cell lysates of wild-type Halobacterium sp. strain NRC-1, gas vesicle-deficient mutants, and purified gas vesicles, after purification of LacZ fusion antibodies on protein A and β-galactosidase affinity columns. Our results show the presence of five new gas vesicle proteins (GvpF, GvpG, GvpJ, GvpL, and GvpM), bringing the total number of proteins identified in the organelles to seven. Two of the new gas vesicle proteins are similar to GvpA (GvpJ and GvpM), and two proteins contain predicted coiled-coil domains (GvpF and GvpL). GvpL exhibited a multiplet ladder on sodium dodecyl sulfate-polyacrylamide gels indicative of oligomerization and self-assembly. We discuss the possible functions of the newly discovered gas vesicle proteins in biogenesis of these unique prokaryotic flotation organelles.

Original languageEnglish (US)
Pages (from-to)3182-3186
Number of pages5
JournalJournal of Bacteriology
Volume186
Issue number10
DOIs
StatePublished - May 1 2004
Externally publishedYes

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Halobacterium
Gases
Proteins
Immunoblotting
Organelles
Galactosidases
Staphylococcal Protein A
Multigene Family
Sodium Dodecyl Sulfate
Immune Sera
gas vesicle protein
Genome
Rabbits
Amino Acids
Antibodies
Genes

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Complexity of Gas Vesicle Biogenesis in Halobacterium sp. Strain NRC-1 : Identification of Five New Proteins. / Shukla, Hem; DasSarma, Shiladitya.

In: Journal of Bacteriology, Vol. 186, No. 10, 01.05.2004, p. 3182-3186.

Research output: Contribution to journalArticle

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