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Ureaplasma parvum infection alters filamin A dynamics in host cells.

Allam AB, Alvarez S, Brown MB, Reyes L - BMC Infect. Dis. (2011)

Bottom Line: In the BPH-1 model, we confirmed that U. parvum perturbed the regulation of filamin A.Specifically, infected BPH-1 cells exhibited a significant increase in filamin A phosphorylated at serine 2152 (P ≤ 0.01), which correlated with impaired proteolysis of the protein and its normal intracellular distribution.Phosphorylation of filamin A occurs in response to various cell signaling cascades that regulate cell motility, differentiation, apoptosis and inflammation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Infectious Disease & Pathology, College of Veterinary Medicine, University of Florida, Gainesville, FL, USA.

ABSTRACT

Background: Ureaplasmas are among the most common bacteria isolated from the human urogenital tract. Ureaplasmas can produce asymptomatic infections or disease characterized by an exaggerated inflammatory response. Most investigations have focused on elucidating the pathogenic potential of Ureaplasma species, but little attention has been paid to understanding the mechanisms by which these organisms are capable of establishing asymptomatic infection.

Methods: We employed differential proteome profiling of bladder tissues from rats experimentally infected with U. parvum in order to identify host cell processes perturbed by colonization with the microbe. Tissues were grouped into four categories: sham inoculated controls, animals that spontaneously cleared infection, asymptomatic urinary tract infection (UTI), and complicated UTI. One protein that was perturbed by infection (filamin A) was used to further elucidate the mechanism of U. parvum-induced disruption in human benign prostate cells (BPH-1). BPH-1 cells were evaluated by confocal microscopy, immunoblotting and ELISA.

Results: Bladder tissue from animals actively colonized with U. parvum displayed significant alterations in actin binding proteins (profilin 1, vinculin, α actinin, and filamin A) that regulate both actin polymerization and cell cytoskeletal function pertaining to focal adhesion formation and signal transduction (Fisher's exact test, P < 0.004; ANOVA, P < 0.02). This phenomenon was independent of clinical profile (asymptomatic vs. complicated UTI). We selected filamin A as a target for additional studies. In the BPH-1 model, we confirmed that U. parvum perturbed the regulation of filamin A. Specifically, infected BPH-1 cells exhibited a significant increase in filamin A phosphorylated at serine 2152 (P ≤ 0.01), which correlated with impaired proteolysis of the protein and its normal intracellular distribution.

Conclusion: Filamin A dynamics were perturbed in both models of infection. Phosphorylation of filamin A occurs in response to various cell signaling cascades that regulate cell motility, differentiation, apoptosis and inflammation. Thus, this phenomenon may be a useful molecular marker for identifying the specific host cell pathways that are perturbed during U. parvum infection.

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Proteome profiling of F344 rat tissues inoculated with sterile broth or U. parvum. Panels A and B represent the percent of proteins assigned to each biological function group. Gene ontology designations were obtained from the Uniprot/Swissprot Database. Protein ratios of each specific protein are from UTI and Struvite groups divided by the Negative group (n = 3). Graph A shows the distribution of protein ratios that exhibited a significant difference by Pro Group™ algorithm (P < 0.05). **Biological function categories that were determined to be significantly different by enrichment analysis. Graph B shows the distribution of protein ratios that were not significantly different between Negative group and groups that were culture positive for U. parvum (UTI and Struvite). Panel C is a hierarchical cluster of standardized least squares means that were significantly different (P < 0.02) among Negative, Struvite, and UTI groups. Ratios generated with the ProGroup™ algorithm were analyzed by ANOVA (n = 3) with a false discovery rate α = 0.05.
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Figure 1: Proteome profiling of F344 rat tissues inoculated with sterile broth or U. parvum. Panels A and B represent the percent of proteins assigned to each biological function group. Gene ontology designations were obtained from the Uniprot/Swissprot Database. Protein ratios of each specific protein are from UTI and Struvite groups divided by the Negative group (n = 3). Graph A shows the distribution of protein ratios that exhibited a significant difference by Pro Group™ algorithm (P < 0.05). **Biological function categories that were determined to be significantly different by enrichment analysis. Graph B shows the distribution of protein ratios that were not significantly different between Negative group and groups that were culture positive for U. parvum (UTI and Struvite). Panel C is a hierarchical cluster of standardized least squares means that were significantly different (P < 0.02) among Negative, Struvite, and UTI groups. Ratios generated with the ProGroup™ algorithm were analyzed by ANOVA (n = 3) with a false discovery rate α = 0.05.

Mentions: Two approaches, enrichment analysis and ANOVA, were used to identify proteins that displayed the same response to U. parvum infection regardless of the clinical profile (UTI or Struvite). In the first approach, enrichment analysis was performed on protein ratios that were generated by comparing the Negative group to animals that were still actively infected with U. parvum (both UTI and Struvite groups). Only proteins that were identified in all three independent iTRAQ experiments and that also exhibited a common effect among animals actively infected with U. parvum were used for this analysis. Using these criteria, 28 of 84 proteins exhibited both a significant (P < 0.05) and a common effect with U. parvum colonization (listed in Additional file 1, Table S1). The distribution of these proteins according to their biological function is summarized in Figure 1a and 1b. Enrichment analysis revealed that animals colonized with U. parvum exhibited a significant change in proteins that regulate actin polymerization (P < 0.004, with Bonferroni correction). These actin-regulating proteins were profilin 1, filamin A, α actinin, vinculin, spectrin and talin. With the exception of profilin 1, all actin binding proteins were significantly lower in animals colonized with U. parvum (Additional File 1, Table S1).


Ureaplasma parvum infection alters filamin A dynamics in host cells.

Allam AB, Alvarez S, Brown MB, Reyes L - BMC Infect. Dis. (2011)

Proteome profiling of F344 rat tissues inoculated with sterile broth or U. parvum. Panels A and B represent the percent of proteins assigned to each biological function group. Gene ontology designations were obtained from the Uniprot/Swissprot Database. Protein ratios of each specific protein are from UTI and Struvite groups divided by the Negative group (n = 3). Graph A shows the distribution of protein ratios that exhibited a significant difference by Pro Group™ algorithm (P < 0.05). **Biological function categories that were determined to be significantly different by enrichment analysis. Graph B shows the distribution of protein ratios that were not significantly different between Negative group and groups that were culture positive for U. parvum (UTI and Struvite). Panel C is a hierarchical cluster of standardized least squares means that were significantly different (P < 0.02) among Negative, Struvite, and UTI groups. Ratios generated with the ProGroup™ algorithm were analyzed by ANOVA (n = 3) with a false discovery rate α = 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3107797&req=5

Figure 1: Proteome profiling of F344 rat tissues inoculated with sterile broth or U. parvum. Panels A and B represent the percent of proteins assigned to each biological function group. Gene ontology designations were obtained from the Uniprot/Swissprot Database. Protein ratios of each specific protein are from UTI and Struvite groups divided by the Negative group (n = 3). Graph A shows the distribution of protein ratios that exhibited a significant difference by Pro Group™ algorithm (P < 0.05). **Biological function categories that were determined to be significantly different by enrichment analysis. Graph B shows the distribution of protein ratios that were not significantly different between Negative group and groups that were culture positive for U. parvum (UTI and Struvite). Panel C is a hierarchical cluster of standardized least squares means that were significantly different (P < 0.02) among Negative, Struvite, and UTI groups. Ratios generated with the ProGroup™ algorithm were analyzed by ANOVA (n = 3) with a false discovery rate α = 0.05.
Mentions: Two approaches, enrichment analysis and ANOVA, were used to identify proteins that displayed the same response to U. parvum infection regardless of the clinical profile (UTI or Struvite). In the first approach, enrichment analysis was performed on protein ratios that were generated by comparing the Negative group to animals that were still actively infected with U. parvum (both UTI and Struvite groups). Only proteins that were identified in all three independent iTRAQ experiments and that also exhibited a common effect among animals actively infected with U. parvum were used for this analysis. Using these criteria, 28 of 84 proteins exhibited both a significant (P < 0.05) and a common effect with U. parvum colonization (listed in Additional file 1, Table S1). The distribution of these proteins according to their biological function is summarized in Figure 1a and 1b. Enrichment analysis revealed that animals colonized with U. parvum exhibited a significant change in proteins that regulate actin polymerization (P < 0.004, with Bonferroni correction). These actin-regulating proteins were profilin 1, filamin A, α actinin, vinculin, spectrin and talin. With the exception of profilin 1, all actin binding proteins were significantly lower in animals colonized with U. parvum (Additional File 1, Table S1).

Bottom Line: In the BPH-1 model, we confirmed that U. parvum perturbed the regulation of filamin A.Specifically, infected BPH-1 cells exhibited a significant increase in filamin A phosphorylated at serine 2152 (P ≤ 0.01), which correlated with impaired proteolysis of the protein and its normal intracellular distribution.Phosphorylation of filamin A occurs in response to various cell signaling cascades that regulate cell motility, differentiation, apoptosis and inflammation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Infectious Disease & Pathology, College of Veterinary Medicine, University of Florida, Gainesville, FL, USA.

ABSTRACT

Background: Ureaplasmas are among the most common bacteria isolated from the human urogenital tract. Ureaplasmas can produce asymptomatic infections or disease characterized by an exaggerated inflammatory response. Most investigations have focused on elucidating the pathogenic potential of Ureaplasma species, but little attention has been paid to understanding the mechanisms by which these organisms are capable of establishing asymptomatic infection.

Methods: We employed differential proteome profiling of bladder tissues from rats experimentally infected with U. parvum in order to identify host cell processes perturbed by colonization with the microbe. Tissues were grouped into four categories: sham inoculated controls, animals that spontaneously cleared infection, asymptomatic urinary tract infection (UTI), and complicated UTI. One protein that was perturbed by infection (filamin A) was used to further elucidate the mechanism of U. parvum-induced disruption in human benign prostate cells (BPH-1). BPH-1 cells were evaluated by confocal microscopy, immunoblotting and ELISA.

Results: Bladder tissue from animals actively colonized with U. parvum displayed significant alterations in actin binding proteins (profilin 1, vinculin, α actinin, and filamin A) that regulate both actin polymerization and cell cytoskeletal function pertaining to focal adhesion formation and signal transduction (Fisher's exact test, P < 0.004; ANOVA, P < 0.02). This phenomenon was independent of clinical profile (asymptomatic vs. complicated UTI). We selected filamin A as a target for additional studies. In the BPH-1 model, we confirmed that U. parvum perturbed the regulation of filamin A. Specifically, infected BPH-1 cells exhibited a significant increase in filamin A phosphorylated at serine 2152 (P ≤ 0.01), which correlated with impaired proteolysis of the protein and its normal intracellular distribution.

Conclusion: Filamin A dynamics were perturbed in both models of infection. Phosphorylation of filamin A occurs in response to various cell signaling cascades that regulate cell motility, differentiation, apoptosis and inflammation. Thus, this phenomenon may be a useful molecular marker for identifying the specific host cell pathways that are perturbed during U. parvum infection.

Show MeSH
Related in: MedlinePlus