Faculty Opinions recommendation of Identification of genes of VSH-1, a prophage-like gene transfer agent of Brachyspira hyodysenteriae.

ABSTRACT The Brachyspira hyodysenteriae B204 genome sequence revealed three VSH-1 tail genes, hvp31, hvp60, and hvp37, in a 3.6-kb cluster. The location and transcription direction of these genes relative to those of the previously described VSH-1 16.3-kb gene operon indicate that the gene transfer agent VSH-1 has a noncontiguous, divided genome.

Identification of Genes of VSH-1, a Prophage-Like Gene Transfer Agent of Brachyspira hyodysenteriae

Journal of Bacteriology ◽

10.1128/jb.187.17.5885-5892.2005 ◽

2005 ◽

Vol 187 (17) ◽

pp. 5885-5892 ◽

Cited By ~ 43

Author(s):

Eric G. Matson ◽

M. Greg Thompson ◽

Samuel B. Humphrey ◽

Richard L. Zuerner ◽

Thad B. Stanton

Keyword(s):

Amino Acid ◽

Gene Transfer ◽

Rhodobacter Capsulatus ◽

Bacterial Species ◽

Polyacrylamide Gel Electrophoresis ◽

Bacterial Genome ◽

Sodium Dodecyl ◽

Amino Acid Sequences ◽

ABSTRACT VSH-1 is a mitomycin C-inducible prophage of the anaerobic spirochete Brachyspira hyodysenteriae. Purified VSH-1 virions are noninfectious, contain random 7.5-kb fragments of the bacterial genome, and mediate generalized transduction of B. hyodysenteriae cells. In order to identify and sequence genes of this novel gene transfer agent (GTA), proteins associated either with VSH-1 capsids or with tails were purified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The N-terminal amino acid sequences of 11 proteins were determined. Degenerate PCR primers were designed from the amino acid sequences and used to amplify several VSH-1 genes from B. hyodysenteriae strain B204 DNA. A λ clone library of B. hyodysenteriae B204 DNA was subsequently screened by Southern hybridization methods and used to identify and sequence overlapping DNA inserts containing additional VSH-1 genes. VSH-1 genes spanned 16.3 kb of the B. hyodysenteriae chromosome and were flanked by bacterial genes. VSH-1 identified genes and unidentified, intervening open reading frames were consecutively organized in head (seven genes), tail (seven genes), and lysis (four genes) clusters in the same transcriptional direction. Putative lysis genes encoding endolysin (Lys) and holin proteins were identified from sequence and structural similarities of their translated protein products with GenBank bacteriophage proteins. Recombinant Lys protein hydrolyzed peptidoglycan purified from B. hyodysenteriae cells. The identified VSH-1 genes exceed the DNA capacity of VSH-1 virions and do not encode traditional bacteriophage early functions involved in DNA replication. These genome properties explain the noninfectious nature of VSH-1 virions and further confirm its resemblance to known prophage-like, GTAs of other bacterial species, such as the GTA from Rhodobacter capsulatus. The identification of VSH-1 genes will enable analysis of the regulation of this GTA and should facilitate investigations of VSH-1-like prophages from other Brachyspira species.

Induction and transcription of VSH-1, a prophage-like gene transfer agent of Brachyspira hyodysenteriae

Anaerobe ◽

10.1016/j.anaerobe.2007.04.004 ◽

2007 ◽

Vol 13 (2) ◽

pp. 89-97 ◽

Cited By ~ 8

Author(s):

Eric G. Matson ◽

Richard L. Zuerner ◽

Thad B. Stanton

Keyword(s):

Gene Transfer ◽

Regulation and release of VSH-1, a prophage and gene transfer agent of Brachyspira hyodysenteriae

10.31274/rtd-180813-12389 ◽

2004 ◽

Author(s):

Eric Glenn Matson

Keyword(s):

Gene Transfer ◽

Collateral Effects of Antibiotics: Carbadox and Metronidazole Induce VSH-1 and Facilitate Gene Transfer among Brachyspira hyodysenteriae Strains

Applied and Environmental Microbiology ◽

10.1128/aem.00189-08 ◽

2008 ◽

Vol 74 (10) ◽

pp. 2950-2956 ◽

Cited By ~ 33

Author(s):

Thaddeus B. Stanton ◽

Samuel B. Humphrey ◽

Vijay K. Sharma ◽

Richard L. Zuerner

Keyword(s):

Gene Transfer ◽

Mitomycin C ◽

Intestinal Bacteria ◽

Etiologic Agent ◽

Chloramphenicol Resistance ◽

Transfer Genes ◽

Collateral Effects ◽

Gene Transfer Agent ◽

Head Protein

ABSTRACT Brachyspira hyodysenteriae is an anaerobic spirochete and the etiologic agent of swine dysentery. The genome of this spirochete contains a mitomycin C-inducible, prophage-like gene transfer agent designated VSH-1. VSH-1 particles package random 7.5-kb fragments of the B. hyodysenteriae genome and transfer genes between B. hyodysenteriae cells. The chemicals and conditions inducing VSH-1 production are largely unknown. Antibiotics used in swine management and stressors inducing traditional prophages might induce VSH-1 and thereby stimulate lateral gene transfer between B. hyodysenteriae cells. In these studies, VSH-1 induction was initially detected by a quantitative real-time reverse transcriptase PCR assay evaluating increased transcription of hvp38 (VSH-1 head protein gene). VSH-1 induction was confirmed by detecting VSH-1-associated 7.5-kb DNA and VSH-1 particles in B. hyodysenteriae cultures. Nine antibiotics (chlortetracycline, lincomycin, tylosin, tiamulin, virginiamycin, ampicillin, ceftriaxone, vancomycin, and florfenicol) at concentrations affecting B. hyodysenteriae growth did not induce VSH-1 production. By contrast, VSH-1 was detected in B. hyodysenteriae cultures treated with mitomycin C (10 μg/ml), carbadox (0.5 μg/ml), metronidazole (0.5 μg/ml), and H2O2 (300 μM). Carbadox- and metronidazole-induced VSH-1 particles transmitted tylosin and chloramphenicol resistance determinants between B. hyodysenteriae strains. The results of these studies suggest that certain antibiotics may induce the production of prophage or prophage-like elements by intestinal bacteria and thereby impact intestinal microbial ecology.

Occurrence and Expression of Gene Transfer Agent Genes in Marine Bacterioplankton

Applied and Environmental Microbiology ◽

10.1128/aem.02129-07 ◽

2008 ◽

Vol 74 (10) ◽

pp. 2933-2939 ◽

Cited By ~ 69

Author(s):

Erin J. Biers ◽

Kui Wang ◽

Catherine Pennington ◽

Robert Belas ◽

Feng Chen ◽

...

Keyword(s):

Gene Transfer ◽

Sequence Data ◽

Genome Sequences ◽

Metagenomic Sequence ◽

Marine Bacterioplankton ◽

Extracellular Release ◽

Gene Transfer Agent ◽

Metagenomic Sequence Data ◽

Search For Homologs

ABSTRACT Genes with homology to the transduction-like gene transfer agent (GTA) were observed in genome sequences of three cultured members of the marine Roseobacter clade. A broader search for homologs for this host-controlled virus-like gene transfer system identified likely GTA systems in cultured Alphaproteobacteria, and particularly in marine bacterioplankton representatives. Expression of GTA genes and extracellular release of GTA particles (∼50 to 70 nm) was demonstrated experimentally for the Roseobacter clade member Silicibacter pomeroyi DSS-3, and intraspecific gene transfer was documented. GTA homologs are surprisingly infrequent in marine metagenomic sequence data, however, and the role of this lateral gene transfer mechanism in ocean bacterioplankton communities remains unclear.

Gene Transfer Agent Promotes Evolvability within the Fittest Subpopulation of a Bacterial Pathogen

Cell Systems ◽

10.1016/j.cels.2017.05.011 ◽

2017 ◽

Vol 4 (6) ◽

pp. 611-621.e6 ◽

Cited By ~ 18

Author(s):

Maxime Québatte ◽

Matthias Christen ◽

Alexander Harms ◽

Jonas Körner ◽

Beat Christen ◽

...

Keyword(s):

Gene Transfer ◽

Bacterial Pathogen ◽

Environmental Factors Influencing Gene Transfer Agent (GTA) Mediated Transduction in the Subtropical Ocean

PLoS ONE ◽

10.1371/journal.pone.0043506 ◽

2012 ◽

Vol 7 (8) ◽

pp. e43506 ◽

Cited By ~ 23

Author(s):

Lauren D. McDaniel ◽

Elizabeth C. Young ◽

Kimberly B. Ritchie ◽

John H. Paul

Keyword(s):

Gene Transfer ◽

Environmental Factors ◽

Factors Influencing ◽

The Protease ClpXP and the PAS Domain Protein DivL Regulate CtrA and Gene Transfer Agent Production inRhodobacter capsulatus

Applied and Environmental Microbiology ◽

10.1128/aem.00275-18 ◽

2018 ◽

Vol 84 (11) ◽

Cited By ~ 9

Author(s):

Alexander B. Westbye ◽

Lukas Kater ◽

Christina Wiesmann ◽

Hao Ding ◽

Calvin K. Yip ◽

...

Keyword(s):

Gene Transfer ◽

Horizontal Gene Transfer ◽

Rhodobacter Capsulatus ◽

Caulobacter Crescentus ◽

Response Regulator ◽

Pas Domain ◽

Content Type ◽

Regulatory Systems ◽

Gene Transfer Agent ◽

Domain Protein

ABSTRACTSeveral members of theRhodobacterales(Alphaproteobacteria) produce a conserved horizontal gene transfer vector, called the gene transfer agent (GTA), that appears to have evolved from a bacteriophage. The model system used to study GTA biology is theRhodobacter capsulatusGTA (RcGTA), a small, tailed bacteriophage-like particle produced by a subset of the cells in a culture. The response regulator CtrA is conserved in theAlphaproteobacteriaand is an essential regulator of RcGTA production: it controls the production and maturation of the RcGTA particle and RcGTA release from cells. CtrA also controls the natural transformation-like system required for cells to receive RcGTA-donated DNA. Here, we report that dysregulation of the CckA-ChpT-CtrA phosphorelay either by the loss of the PAS domain protein DivL or by substitution of the autophosphorylation residue of the hybrid histidine kinase CckA decreased CtrA phosphorylation and greatly increased RcGTA protein production inR. capsulatus. We show that the loss of the ClpXP protease or the three C-terminal residues of CtrA results in increased CtrA levels inR. capsulatusand identify ClpX(P) to be essential for the maturation of RcGTA particles. Furthermore, we show that CtrA phosphorylation is important for head spike production. Our results provide novel insight into the regulation of CtrA and GTAs in theRhodobacterales.IMPORTANCEMembers of theRhodobacteralesare abundant in ocean and freshwater environments. The conserved GTA produced by manyRhodobacteralesmay have an important role in horizontal gene transfer (HGT) in aquatic environments and provide a significant contribution to their adaptation. GTA production is controlled by bacterial regulatory systems, including the conserved CckA-ChpT-CtrA phosphorelay; however, several questions about GTA regulation remain. Our identification that a short DivL homologue and ClpXP regulate CtrA inR. capsulatusextends the model of CtrA regulation fromCaulobacter crescentusto a member of theRhodobacterales. We found that the magnitude of RcGTA production greatly depends on DivL and CckA kinase activity, adding yet another layer of regulatory complexity to RcGTA. RcGTA is known to undergo CckA-dependent maturation, and we extend the understanding of this process by showing that the ClpX chaperone is required for formation of tailed, DNA-containing particles.