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- Title
Proteomic Analysis of the Action of the Mycobacterium ulcerans Toxin Mycolactone: Targeting Host Cells Cytoskeleton and Collagen.
- Authors
Gama, José B.; Ohlmeier, Steffen; Martins, Teresa G.; Fraga, Alexandra G.; Sampaio-Marques, Belém; Carvalho, Maria A.; Proença, Fernanda; Silva, Manuel T.; Pedrosa, Jorge; Ludovico, Paula
- Abstract
Buruli ulcer (BU) is a neglected tropical disease caused by Mycobacterium ulcerans. The tissue damage characteristic of BU lesions is known to be driven by the secretion of the potent lipidic exotoxin mycolactone. However, the molecular action of mycolactone on host cell biology mediating cytopathogenesis is not fully understood. Here we applied two-dimensional electrophoresis (2-DE) to identify the mechanisms of mycolactone's cellular action in the L929 mouse fibroblast proteome. This revealed 20 changed spots corresponding to 18 proteins which were clustered mainly into cytoskeleton-related proteins (Dync1i2, Cfl1, Crmp2, Actg1, Stmn1) and collagen biosynthesis enzymes (Plod1, Plod3, P4ha1). In line with cytoskeleton conformational disarrangements that are observed by immunofluorescence, we found several regulators and constituents of both actin- and tubulin-cytoskeleton affected upon exposure to the toxin, providing a novel molecular basis for the effect of mycolactone. Consistent with these cytoskeleton-related alterations, accumulation of autophagosomes as well as an increased protein ubiquitination were observed in mycolactone-treated cells. In vivo analyses in a BU mouse model revealed mycolactone-dependent structural changes in collagen upon infection with M. ulcerans, associated with the reduction of dermal collagen content, which is in line with our proteomic finding of mycolactone-induced down-regulation of several collagen biosynthesis enzymes. Our results unveil the mechanisms of mycolactone-induced molecular cytopathogenesis on exposed host cells, with the toxin compromising cell structure and homeostasis by inducing cytoskeleton alterations, as well as disrupting tissue structure, by impairing the extracellular matrix biosynthesis. Author Summary: Buruli Ulcer (BU) is a neglected tropical disease caused by Mycobacterium ulcerans infection. It has been recognized for many years that BU pathogenesis is mediated by the potent exotoxin mycolactone; however, the molecular action of this toxin on the host cell biology that drives its pathogenesis is not fully understood. Here we present a proteomic-based study that explores the molecular action of mycolactone on host cells biology. Our results provide further molecular evidence for the cytoskeleton-disarrangement induced by mycolactone, and unveil its impact on cytoskeleton-dependent cellular functions. Moreover, we extend the field of action of this toxin to the biosynthesis of collagen, implicating mycolactone on the decrease of dermal collagen found on BU lesions. Given the dependence of M. ulcerans virulence on its toxin, these findings on mycolactone's molecular action on host cells and tissues are of major importance for the understanding of BU pathogenesis.
- Subjects
EXOTOXIN; BACTERIAL toxins; PROTEOMICS; CYTOLOGY; TUBULINS; COLLAGEN; TOXINS; CYTOSKELETON
- Publication
PLoS Neglected Tropical Diseases, 2014, Vol 8, Issue 8, p1
- ISSN
1935-2727
- Publication type
Article
- DOI
10.1371/journal.pntd.0003066