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Altered Proteome of Burkholderia pseudomallei Colony Variants Induced by Exposure to Human Lung Epithelial Cells.

Al-Maleki AR, Mariappan V, Vellasamy KM, Tay ST, Vadivelu J - PLoS ONE (2015)

Bottom Line: Burkholderia pseudomallei primary diagnostic cultures demonstrate colony morphology variation associated with expression of virulence and adaptation proteins.Additionally, post-exposure expression profiles of both variants were compared with pre-exposure.Protein profiles differences post-exposure provide insights into association between morphotypic and phenotypic characteristics of colony variants, strengthening the role of B. pseudomallei morphotypes in pathogenesis of melioidosis.

View Article: PubMed Central - PubMed

Affiliation: Tropical Infectious Disease Research and Education Center (TIDREC), Department of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.

ABSTRACT
Burkholderia pseudomallei primary diagnostic cultures demonstrate colony morphology variation associated with expression of virulence and adaptation proteins. This study aims to examine the ability of B. pseudomallei colony variants (wild type [WT] and small colony variant [SCV]) to survive and replicate intracellularly in A549 cells and to identify the alterations in the protein expression of these variants, post-exposure to the A549 cells. Intracellular survival and cytotoxicity assays were performed followed by proteomics analysis using two-dimensional gel electrophoresis. B. pseudomallei SCV survive longer than the WT. During post-exposure, among 259 and 260 protein spots of SCV and WT, respectively, 19 were differentially expressed. Among SCV post-exposure up-regulated proteins, glyceraldehyde 3-phosphate dehydrogenase, fructose-bisphosphate aldolase (CbbA) and betaine aldehyde dehydrogenase were associated with adhesion and virulence. Among the down-regulated proteins, enolase (Eno) is implicated in adhesion and virulence. Additionally, post-exposure expression profiles of both variants were compared with pre-exposure. In WT pre- vs post-exposure, 36 proteins were differentially expressed. Of the up-regulated proteins, translocator protein, Eno, nucleoside diphosphate kinase (Ndk), ferritin Dps-family DNA binding protein and peptidyl-prolyl cis-trans isomerase B were implicated in invasion and virulence. In SCV pre- vs post-exposure, 27 proteins were differentially expressed. Among the up-regulated proteins, flagellin, Eno, CbbA, Ndk and phenylacetate-coenzyme A ligase have similarly been implicated in adhesion, invasion. Protein profiles differences post-exposure provide insights into association between morphotypic and phenotypic characteristics of colony variants, strengthening the role of B. pseudomallei morphotypes in pathogenesis of melioidosis.

No MeSH data available.


Related in: MedlinePlus

Protein categories and the subcellular localisation of Burkholderia pseudomallei WT (pre-exposure vs post-exposure to A549 cells).Functional protein categories were predicted using COG, while subcellular localisation was predicted using PSORT. Panels A and B refer to functions and cellular location of the up-regulated proteins, respectively. Panels C and D refer to functions and cellular location of the down-regulated proteins, respectively.
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pone.0127398.g007: Protein categories and the subcellular localisation of Burkholderia pseudomallei WT (pre-exposure vs post-exposure to A549 cells).Functional protein categories were predicted using COG, while subcellular localisation was predicted using PSORT. Panels A and B refer to functions and cellular location of the up-regulated proteins, respectively. Panels C and D refer to functions and cellular location of the down-regulated proteins, respectively.

Mentions: Among the up-regulated proteins that were detected in the SCV post-exposure comparing with SCV pre-exposure to A549 cells, 10 proteins (Eno, CbbA, Fpr, BPSL2288, HmpA, AhcY, PaaA, DapD, ArcB and Ndk) were implicated in metabolism of the bacteria and one (FliC) in cellular processes (cell motility) (Table 4; Fig 7A). However, localisation prediction revealed that 90.9% were of cytoplasmic and 9.1% of extracellular (Fig 7B). Whereas, among the down-regulated proteins, seven (BkdA2, ScoB, ScoA, PhbA, Adk, ArgT, and BPSL1167) were involved in metabolism and three in cellular processes (BPSS2288, GroEL1 (n = 5) and BPSL2748) (Fig 7C). Of the 10 down-regulated proteins, 80% were of cytoplasmic origin, 10% of periplasmic and 1% of multiple localisation sites (Fig 7D).


Altered Proteome of Burkholderia pseudomallei Colony Variants Induced by Exposure to Human Lung Epithelial Cells.

Al-Maleki AR, Mariappan V, Vellasamy KM, Tay ST, Vadivelu J - PLoS ONE (2015)

Protein categories and the subcellular localisation of Burkholderia pseudomallei WT (pre-exposure vs post-exposure to A549 cells).Functional protein categories were predicted using COG, while subcellular localisation was predicted using PSORT. Panels A and B refer to functions and cellular location of the up-regulated proteins, respectively. Panels C and D refer to functions and cellular location of the down-regulated proteins, respectively.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4440636&req=5

pone.0127398.g007: Protein categories and the subcellular localisation of Burkholderia pseudomallei WT (pre-exposure vs post-exposure to A549 cells).Functional protein categories were predicted using COG, while subcellular localisation was predicted using PSORT. Panels A and B refer to functions and cellular location of the up-regulated proteins, respectively. Panels C and D refer to functions and cellular location of the down-regulated proteins, respectively.
Mentions: Among the up-regulated proteins that were detected in the SCV post-exposure comparing with SCV pre-exposure to A549 cells, 10 proteins (Eno, CbbA, Fpr, BPSL2288, HmpA, AhcY, PaaA, DapD, ArcB and Ndk) were implicated in metabolism of the bacteria and one (FliC) in cellular processes (cell motility) (Table 4; Fig 7A). However, localisation prediction revealed that 90.9% were of cytoplasmic and 9.1% of extracellular (Fig 7B). Whereas, among the down-regulated proteins, seven (BkdA2, ScoB, ScoA, PhbA, Adk, ArgT, and BPSL1167) were involved in metabolism and three in cellular processes (BPSS2288, GroEL1 (n = 5) and BPSL2748) (Fig 7C). Of the 10 down-regulated proteins, 80% were of cytoplasmic origin, 10% of periplasmic and 1% of multiple localisation sites (Fig 7D).

Bottom Line: Burkholderia pseudomallei primary diagnostic cultures demonstrate colony morphology variation associated with expression of virulence and adaptation proteins.Additionally, post-exposure expression profiles of both variants were compared with pre-exposure.Protein profiles differences post-exposure provide insights into association between morphotypic and phenotypic characteristics of colony variants, strengthening the role of B. pseudomallei morphotypes in pathogenesis of melioidosis.

View Article: PubMed Central - PubMed

Affiliation: Tropical Infectious Disease Research and Education Center (TIDREC), Department of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.

ABSTRACT
Burkholderia pseudomallei primary diagnostic cultures demonstrate colony morphology variation associated with expression of virulence and adaptation proteins. This study aims to examine the ability of B. pseudomallei colony variants (wild type [WT] and small colony variant [SCV]) to survive and replicate intracellularly in A549 cells and to identify the alterations in the protein expression of these variants, post-exposure to the A549 cells. Intracellular survival and cytotoxicity assays were performed followed by proteomics analysis using two-dimensional gel electrophoresis. B. pseudomallei SCV survive longer than the WT. During post-exposure, among 259 and 260 protein spots of SCV and WT, respectively, 19 were differentially expressed. Among SCV post-exposure up-regulated proteins, glyceraldehyde 3-phosphate dehydrogenase, fructose-bisphosphate aldolase (CbbA) and betaine aldehyde dehydrogenase were associated with adhesion and virulence. Among the down-regulated proteins, enolase (Eno) is implicated in adhesion and virulence. Additionally, post-exposure expression profiles of both variants were compared with pre-exposure. In WT pre- vs post-exposure, 36 proteins were differentially expressed. Of the up-regulated proteins, translocator protein, Eno, nucleoside diphosphate kinase (Ndk), ferritin Dps-family DNA binding protein and peptidyl-prolyl cis-trans isomerase B were implicated in invasion and virulence. In SCV pre- vs post-exposure, 27 proteins were differentially expressed. Among the up-regulated proteins, flagellin, Eno, CbbA, Ndk and phenylacetate-coenzyme A ligase have similarly been implicated in adhesion, invasion. Protein profiles differences post-exposure provide insights into association between morphotypic and phenotypic characteristics of colony variants, strengthening the role of B. pseudomallei morphotypes in pathogenesis of melioidosis.

No MeSH data available.


Related in: MedlinePlus