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An insertion sequence-dependent plasmid rearrangement in Aeromonas salmonicida causes the loss of the type three secretion system.

Tanaka KH, Dallaire-Dufresne S, Daher RK, Frenette M, Charette SJ - PLoS ONE (2012)

Bottom Line: Nine out of the 26 strains had a positive PCR result, suggesting that the rearrangement in these strains were IS-dependent.Our results suggested that pAsa5 rearrangements involve IS11.This is the first study showing that ISs are involved in plasmid instability in A. salmonicida.

View Article: PubMed Central - PubMed

Affiliation: Institut de biologie intégrative et des systèmes, Pavillon Charles-Eugène-Marchand, Université Laval, Quebec City, Quebec, Canada.

ABSTRACT
Aeromonas salmonicida, a bacterial fish pathogen, possesses a functional Type Three Secretion System (TTSS), which is essential for its virulence. The genes for this system are mainly located in a single region of the large pAsa5 plasmid. Bacteria lose the TTSS region from this plasmid through rearrangements when grown in stressful growth conditions. The A. salmonicida genome is rich in insertion sequences (ISs), which are mobile DNA elements that can cause DNA rearrangements in other bacterial species. pAsa5 possesses numerous ISs. Three IS11s from the IS256 family encircle the rearranged regions. To confirm that these IS11s are involved in pAsa5 rearrangements, 26 strains derived from strain A449 and two Canadian isolates (01-B526 and 01-B516) with a pAsa5 rearrangement were tested using a PCR approach to determine whether the rearrangements were the result of an IS11-dependent process. Nine out of the 26 strains had a positive PCR result, suggesting that the rearrangement in these strains were IS-dependent. The PCR analysis showed that all the rearrangements in the A449-derived strains were IS11-dependent process while the rearrangements in 01-B526 and 01-B516 could only be partially coupled to the action of IS11. Unidentified elements that affect IS-dependent rearrangements may be present in 01-B526 and 01-B516. Our results suggested that pAsa5 rearrangements involve IS11. This is the first study showing that ISs are involved in plasmid instability in A. salmonicida.

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Related in: MedlinePlus

Growing A. salmonicida in stressful conditions can lead to the loss of pAsal1.A and C. Small plasmids (pAsa1, pAsa2, pAsa3 and pAsal1) from 01-B526 and 01-B516 derived strains were purified and digested using EcoRI leading to the linearization of the plasmids [12]. An agarose gel electrophoresis of the digested plasmids is shown. B and D. DNA lysates of the same strains were tested by PCR with pAsal1 primers (Table 2) to confirm by a second approach the presence or the absence of pAsal1 in these strains. The presence (+) or the absence (−) of a PCR product is indicated.
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pone-0033725-g005: Growing A. salmonicida in stressful conditions can lead to the loss of pAsal1.A and C. Small plasmids (pAsa1, pAsa2, pAsa3 and pAsal1) from 01-B526 and 01-B516 derived strains were purified and digested using EcoRI leading to the linearization of the plasmids [12]. An agarose gel electrophoresis of the digested plasmids is shown. B and D. DNA lysates of the same strains were tested by PCR with pAsal1 primers (Table 2) to confirm by a second approach the presence or the absence of pAsal1 in these strains. The presence (+) or the absence (−) of a PCR product is indicated.

Mentions: see Figure 5.


An insertion sequence-dependent plasmid rearrangement in Aeromonas salmonicida causes the loss of the type three secretion system.

Tanaka KH, Dallaire-Dufresne S, Daher RK, Frenette M, Charette SJ - PLoS ONE (2012)

Growing A. salmonicida in stressful conditions can lead to the loss of pAsal1.A and C. Small plasmids (pAsa1, pAsa2, pAsa3 and pAsal1) from 01-B526 and 01-B516 derived strains were purified and digested using EcoRI leading to the linearization of the plasmids [12]. An agarose gel electrophoresis of the digested plasmids is shown. B and D. DNA lysates of the same strains were tested by PCR with pAsal1 primers (Table 2) to confirm by a second approach the presence or the absence of pAsal1 in these strains. The presence (+) or the absence (−) of a PCR product is indicated.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3303853&req=5

pone-0033725-g005: Growing A. salmonicida in stressful conditions can lead to the loss of pAsal1.A and C. Small plasmids (pAsa1, pAsa2, pAsa3 and pAsal1) from 01-B526 and 01-B516 derived strains were purified and digested using EcoRI leading to the linearization of the plasmids [12]. An agarose gel electrophoresis of the digested plasmids is shown. B and D. DNA lysates of the same strains were tested by PCR with pAsal1 primers (Table 2) to confirm by a second approach the presence or the absence of pAsal1 in these strains. The presence (+) or the absence (−) of a PCR product is indicated.
Mentions: see Figure 5.

Bottom Line: Nine out of the 26 strains had a positive PCR result, suggesting that the rearrangement in these strains were IS-dependent.Our results suggested that pAsa5 rearrangements involve IS11.This is the first study showing that ISs are involved in plasmid instability in A. salmonicida.

View Article: PubMed Central - PubMed

Affiliation: Institut de biologie intégrative et des systèmes, Pavillon Charles-Eugène-Marchand, Université Laval, Quebec City, Quebec, Canada.

ABSTRACT
Aeromonas salmonicida, a bacterial fish pathogen, possesses a functional Type Three Secretion System (TTSS), which is essential for its virulence. The genes for this system are mainly located in a single region of the large pAsa5 plasmid. Bacteria lose the TTSS region from this plasmid through rearrangements when grown in stressful growth conditions. The A. salmonicida genome is rich in insertion sequences (ISs), which are mobile DNA elements that can cause DNA rearrangements in other bacterial species. pAsa5 possesses numerous ISs. Three IS11s from the IS256 family encircle the rearranged regions. To confirm that these IS11s are involved in pAsa5 rearrangements, 26 strains derived from strain A449 and two Canadian isolates (01-B526 and 01-B516) with a pAsa5 rearrangement were tested using a PCR approach to determine whether the rearrangements were the result of an IS11-dependent process. Nine out of the 26 strains had a positive PCR result, suggesting that the rearrangement in these strains were IS-dependent. The PCR analysis showed that all the rearrangements in the A449-derived strains were IS11-dependent process while the rearrangements in 01-B526 and 01-B516 could only be partially coupled to the action of IS11. Unidentified elements that affect IS-dependent rearrangements may be present in 01-B526 and 01-B516. Our results suggested that pAsa5 rearrangements involve IS11. This is the first study showing that ISs are involved in plasmid instability in A. salmonicida.

Show MeSH
Related in: MedlinePlus