A large plasmid from Halobacterium halobium carrying genetic information for gas vacuole formation

doi:10.1016/0147-619X(79)90021-0

Plasmid

Volume 2, Issue 3, July 1979, Pages 377-386

https://doi.org/10.1016/0147-619X(79)90021-0 Get rights and content

Abstract

A large plasmid with a molecular weight of 100 × 10⁶ has been found in Halobacterium halobium which is indistinguishable from the previously described “satellite DNA.” In this halophilic bacterium characteristic properties such as the biosyntheses of gas vacuoles, purple membrane, and ruberin are spontaneously lost at high frequencies. These phenotypic alterations are accompanied by a change in the nucleotide sequence of the plasmid DNA. In plasmids of vacuole-deficient mutants two distinct PstI fragments in the restriction pattern are altered probably by an insertion of 3600 base pairs into the DNA. In revenants which form gas vacuoles the original sequence of the plasmid DNA is restored. This indicates that the presence of the plasmid is related to the gas vacuole formation.

References (15)

D. Gillespie et al.
A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane
J. Mol. Biol
(1965)
C.N. Gordon et al.
High contrast staining of individual nucleic acid molecules
Biochim. Biophys. Acta
(1968)
N. Kleckner et al.
Genetic engineering in vivo using translocatable drug resistance elements
J. Mol. Biol
(1977)
E.M. Southern
Detection of specific sequences among DNA fragments separated by gel electrophoresis
J. Mol. Biol
(1975)
M. Sumper et al.
Biosynthesis of purple membrane: Control of refined synthesis by bacterio-opsin
FEBS Lett
(1976)
H. Bünemann et al.
Base specific fractionation of double stranded DNA: Affinity chromatography on a novel type of absorbant
Nucl. Ac. Res
(1978)
G.G. Hayword
Gel electrophoretic separation of the complementary strands of bacteriophage-DNA
Virology
(1972)

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10 Genetic Analysis in the Domain Archaea
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Methods for genetic analysis of Archaea have not been excluded from these improvements. It seems remarkable that the first report of gene transfer in any member of the domain Archaea, that of conjugation in Hf. volcunii, was only about thirteen years ago. The availability of model systems for genetic analysis of Eukarya and of Bacteria strongly influence the types of analyses that are being undertaken in Archaea today, and are in large part responsible for the rapid progress of the field. However, as has been the case with other aspects of archaeal research, it is important to remember that these are not Bacteria, nor are they Eukarya. The unique nature of the Archaea has forced modification of the classical genetic approaches used in the other two domains. Several of the methods described here may have general utility among other Archaea, in addition to the specific uses described in this chapter.
Genetic analysis of the gas vesicle gene cluster in haloarchaea
1997, FEMS Microbiology Letters
Gas vesicles are buoyant intracellular organelles composed of a rigid proteinaceous membrane surrounding a gas-filled space. Many prokaryotic microorganisms including photosynthetic and heterotrophic bacteria and halophilic and methanogenic archaea produce gas vesicles. In the majority of cases gas vesicles function in providing vertical motility to cells in aquatic environments. Recent genetic analysis of several halophilic archaeal (haloarchaeal) species has shown that a large cluster of genes [gvpMLKJIHGFEDACN(O)] is necessary for gas vesicle formation.
Comparative Restriction Maps of the Archaeal Megaplasmid pHM300 in Different Haloferax mediterranei Strains
1995, Systematic and Applied Microbiology
We have constructed the restriction maps of the circular megaplasmid pHM300 of five different Haloferax nrediterranei strains (ATCC 33500, M2, M4, M6, 1110), and a pigmentary mutant from one of them (M2a), using the endonucleases BamHl, SwaI, and BgIII. Either one-dimensional pulsed-field gel electrophoresis (PFGE) or two-dimensional PFGE (2D-PFGE) were used to resolve the genomic fragments generate by these enzymes. For physical mapping, we combined hybridization with multiple digestion straegies. pHM300 maps were identical for strains ATCC 33500/M10 (315 kb), and M-) (M26) /M4 /1106 (330 kb), respectively, with some regions conserved in both map types. The main difference between them appeared to be an insertion that corresponds roughly to a 7 kb SwaI fragment containing an abnormal density of rare-cutter sites as well as repeated sequences. In M2a, pHM300 has only 145 kb, and most likely suffered a large deletion from its original template: pHM300 from M26.
Halobacterial genes and genomes
1993, New Comprehensive Biochemistry
Halobacterial genomes consist of circular DNAs (deoxyribo nucleic acid) similar in size to those of bacteria, apparently without introns in protein-coding genes, and with genes in operons or clusters sometimes closely resembling their bacterial counterparts. The sequences are quite similar, and all have an open reading frame of about 1 kbp and a set of hexanucleotide repeats that may be involved in the maintenance of the plasmid. Comparison of the patterns revealed by hybridization of repeated sequences to independent but not visibly different single-colony isolates of Halobacterium halobium indicated that these elements are highly mobile. The most highly developed genetic map is that of Escherichia coli K12, and it has recently been augmented with coarse and fine restriction maps. As the most nearly complete genetic map, it can give one clues to structural features, which may be functionally important, especially as some data exist on the effects of chromosomal rearrangements. Transformation with cosmid DNA from the Haloferax volcanii minimal set has been used to map 140 ethylmethanesulfonate-induced mutants requiring one of 14 amino acids, uracil, adenine, or guanine.
Large Deletions in Class III Gas Vesicle-Deficient Mutants of Halobacterium halobium
1993, Systematic and Applied Microbiology
Mutants of Halobacterium halobium deficient in production of gas vesicles (Vac^- and Vac^δ-) arise spontaneously at an extremely high frequency, approximately 1%. In this study, we characterized one class of mutants (class III) with a stable and completely Vac^- phenotype. Restriction mapping and Southern analysis of four class III mutants, SD109, SD112A, SD116, and SD118A, demonstrated that the mutants were deleted for the gas vesicle gene cluster and flanking sequences in the 200-kb gas vesicle plasmid, pNRC 100. Deletions resulted from recombination between pNRC 100 fragments HindIII-C and HindIII-D″ (for SD109, SD112A, and SD116) or HindIII-C and an unidentified sequence (for SD118A). In at least three cases, the deletion junctions mapped precisely to the termini of IS elements, ISH2 (for SD112A) and ISH8 (for SD109 and SD118A). These results suggest that deletion formation is a consequence of intramolecular transposition of IS elements, although a multistep mechanism involving recombination between two elements is also possible. Interestingly, a Vac⁺ revenant (SD142R) of a class III mutant (SD142) was isolated from a sectored colony. Southern analysis indicated that SD142 had a deletion of the gvp gene cluster and that SD142R had regained the gas vesicle gene cluster by an unknown mechanism.
Structure and organization of the gas vesicle gene cluster on the Halobacterium halobium plasmid pNRC100
1991, Gene
Halobacterium halobium strain NRC-1 contains intracellular gas-filled vesicles (GVs) that confer buoyancy to the cells. Cloning of the major GV protein (GvpA)-encoding gene, gvpA, and analysis of GV-deficient mutants (Vac⁻) of H. halobium led to the identification of a region of a 200-kb plasmid, pNRC100, important for GV synthesis. We report here the nucleotide sequence of an 8520-bp region which, including gvpA, contains twelve open reading frames (ORFs) that are organized into two divergent transcription units, gvpAC oriented rightward, and gvpD, E, F, G, H, I, J, K, L, and M located upstream from gvpAC and oriented leftward. Insertions into the gvpA promoter and gvpD and E resulted in the Vac⁻ phenotype. The overall gene organization is highly compact with the end of one ORF overlapping with the beginning of the next in most cases. The gene cluster is bracketed by two ISH8 element copies in inverted orientation, an organization suggestive of a composite transposon. Comparison of predicted amino acid sequences showed homology between GvpA, and the gvpJ and gvpM putative gene products. The putative gvpC gene product contains eight copies of an imperfectly repeated sequence with similarity to repeats in a cyanobacterial GvpC plus a highly acidic C-terminal region not found in the cyanobacterial homologue.

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Plasmid

Abstract

References (15)

A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane

J. Mol. Biol

High contrast staining of individual nucleic acid molecules

Biochim. Biophys. Acta

Genetic engineering in vivo using translocatable drug resistance elements

J. Mol. Biol

Detection of specific sequences among DNA fragments separated by gel electrophoresis

J. Mol. Biol

Biosynthesis of purple membrane: Control of refined synthesis by bacterio-opsin

FEBS Lett

Base specific fractionation of double stranded DNA: Affinity chromatography on a novel type of absorbant

Nucl. Ac. Res

Gel electrophoretic separation of the complementary strands of bacteriophage-DNA

Virology

Cited by (47)

10 Genetic Analysis in the Domain Archaea

Genetic analysis of the gas vesicle gene cluster in haloarchaea

Comparative Restriction Maps of the Archaeal Megaplasmid pHM300 in Different Haloferax mediterranei Strains

Halobacterial genes and genomes

Large Deletions in Class III Gas Vesicle-Deficient Mutants of Halobacterium halobium

Structure and organization of the gas vesicle gene cluster on the Halobacterium halobium plasmid pNRC100

Regular articleA large plasmid from Halobacterium halobium carrying genetic information for gas vacuole formation

Abstract

J. Mol. Biol

Biochim. Biophys. Acta

J. Mol. Biol

J. Mol. Biol

FEBS Lett

Base specific fractionation of double stranded DNA: Affinity chromatography on a novel type of absorbant

Nucl. Ac. Res

Gel electrophoretic separation of the complementary strands of bacteriophage-DNA

Virology

Regular article
A large plasmid from Halobacterium halobium carrying genetic information for gas vacuole formation