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Functional Genomics of the Aeromonas salmonicida Lipopolysaccharide O-Antigen and A-Layer from Typical and Atypical Strains.

Merino S, de Mendoza E, Canals R, Tomás JM - Mar Drugs (2015)

Bottom Line: Between genes of the wbsalmo we found the genes responsible for the biosynthesis and assembly of the S-layer (named A-layer in these strains).Through comparative genomic analysis and in-frame deletions of some of the genes, we concluded that all the A. salmonicida typical and atypical strains, other than A. salmonicida subsp. pectinolytica strains, shared the same wbsalmo and presence of A-layer.A. salmonicida subsp. pectinolytica strains lack wbsalmo and A-layer, two major virulence factors, and this could be the reason they are the only ones not found as fish pathogens.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Faculty of Biology, University of Barcelona, Diagonal 643, Barcelona 08071, Spain. smerino@ub.edu.

ABSTRACT
The A. salmonicida A450 LPS O-antigen, encoded by the wbsalmo gene cluster, is exported through an ABC-2 transporter-dependent pathway. It represents the first example of an O-antigen LPS polysaccharide with three different monosaccharides in their repeating unit assembled by this pathway. Until now, only repeating units with one or two different monosaccharides have been described. Functional genomic analysis of this wbsalmo region is mostly in agreement with the LPS O-antigen structure of acetylated l-rhamnose (Rha), d-glucose (Glc), and 2-amino-2-deoxy-d-mannose (ManN). Between genes of the wbsalmo we found the genes responsible for the biosynthesis and assembly of the S-layer (named A-layer in these strains). Through comparative genomic analysis and in-frame deletions of some of the genes, we concluded that all the A. salmonicida typical and atypical strains, other than A. salmonicida subsp. pectinolytica strains, shared the same wbsalmo and presence of A-layer. A. salmonicida subsp. pectinolytica strains lack wbsalmo and A-layer, two major virulence factors, and this could be the reason they are the only ones not found as fish pathogens.

No MeSH data available.


(A) Outer membrane proteins, and (B) Western blot using antiserum against A-layer protein (anti-VapA protein) of strains: A450 (wild type, lane 1), mutant A450ΔvapA (lane 2), A450ΔvapA complemented with A450 vapA (lane 3), and A450ΔUDP-ep (lane 4). St, molecular weight standard.
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marinedrugs-13-03791-f003: (A) Outer membrane proteins, and (B) Western blot using antiserum against A-layer protein (anti-VapA protein) of strains: A450 (wild type, lane 1), mutant A450ΔvapA (lane 2), A450ΔvapA complemented with A450 vapA (lane 3), and A450ΔUDP-ep (lane 4). St, molecular weight standard.

Mentions: We obtained outer-membrane proteins (OMp) from A450ΔvapA mutant as described in Materials and Methods. Analysis by SDS-PAGE gels showed the lack of the major protein band of approximately 49 kDa compared with the wild type (Figure 3A). This band reacts with specific serum anti-VapA protein in Western blot analysis (Figure 3B). Mutant A450ΔUDP-ep showed no changes in OMp.


Functional Genomics of the Aeromonas salmonicida Lipopolysaccharide O-Antigen and A-Layer from Typical and Atypical Strains.

Merino S, de Mendoza E, Canals R, Tomás JM - Mar Drugs (2015)

(A) Outer membrane proteins, and (B) Western blot using antiserum against A-layer protein (anti-VapA protein) of strains: A450 (wild type, lane 1), mutant A450ΔvapA (lane 2), A450ΔvapA complemented with A450 vapA (lane 3), and A450ΔUDP-ep (lane 4). St, molecular weight standard.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4483657&req=5

marinedrugs-13-03791-f003: (A) Outer membrane proteins, and (B) Western blot using antiserum against A-layer protein (anti-VapA protein) of strains: A450 (wild type, lane 1), mutant A450ΔvapA (lane 2), A450ΔvapA complemented with A450 vapA (lane 3), and A450ΔUDP-ep (lane 4). St, molecular weight standard.
Mentions: We obtained outer-membrane proteins (OMp) from A450ΔvapA mutant as described in Materials and Methods. Analysis by SDS-PAGE gels showed the lack of the major protein band of approximately 49 kDa compared with the wild type (Figure 3A). This band reacts with specific serum anti-VapA protein in Western blot analysis (Figure 3B). Mutant A450ΔUDP-ep showed no changes in OMp.

Bottom Line: Between genes of the wbsalmo we found the genes responsible for the biosynthesis and assembly of the S-layer (named A-layer in these strains).Through comparative genomic analysis and in-frame deletions of some of the genes, we concluded that all the A. salmonicida typical and atypical strains, other than A. salmonicida subsp. pectinolytica strains, shared the same wbsalmo and presence of A-layer.A. salmonicida subsp. pectinolytica strains lack wbsalmo and A-layer, two major virulence factors, and this could be the reason they are the only ones not found as fish pathogens.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Faculty of Biology, University of Barcelona, Diagonal 643, Barcelona 08071, Spain. smerino@ub.edu.

ABSTRACT
The A. salmonicida A450 LPS O-antigen, encoded by the wbsalmo gene cluster, is exported through an ABC-2 transporter-dependent pathway. It represents the first example of an O-antigen LPS polysaccharide with three different monosaccharides in their repeating unit assembled by this pathway. Until now, only repeating units with one or two different monosaccharides have been described. Functional genomic analysis of this wbsalmo region is mostly in agreement with the LPS O-antigen structure of acetylated l-rhamnose (Rha), d-glucose (Glc), and 2-amino-2-deoxy-d-mannose (ManN). Between genes of the wbsalmo we found the genes responsible for the biosynthesis and assembly of the S-layer (named A-layer in these strains). Through comparative genomic analysis and in-frame deletions of some of the genes, we concluded that all the A. salmonicida typical and atypical strains, other than A. salmonicida subsp. pectinolytica strains, shared the same wbsalmo and presence of A-layer. A. salmonicida subsp. pectinolytica strains lack wbsalmo and A-layer, two major virulence factors, and this could be the reason they are the only ones not found as fish pathogens.

No MeSH data available.