Use of holograph optical trap: image system to study the adhesin function of Drechmeria coniospora 14-3-3 protein in Caenorhabditis elegans

Abstract

The interaction fungal-cell is essential factor to establish the pathogenesis. The molecules involved in fungal adhesion are known as adhesins and until now few adhesins have been identified from fungal pathogens. For Paracoccidioides species, the glycoprotein of 43 kDa, enolase, malate synthase, glyceraldehyde-3-phosphate dehydrogenase, triosephosphate isomerase has been described. Another protein of 30 kDa which belongs to 14-3-3 protein family is highlighting as an adhesion of this fungus and may have an important role in paracoccidioidomycosis. 14-3-3 proteins are present in all eukaryotes, highly conserved and have the ability to bind to other proteins in eukaryotes resulting in both activation and inhibition of biological processes; therefore 14-3-3 has important and complex participation in biological processes. In Paracoccidioides spp. 14-3-3 protein has been described as an adhesin and expression was higher in isolates with high adhesion rate. Despite many years of study of paracoccidioidomycosis, 14-3-3 protein do not have its action elucidated in the host, in addition, because ethical problems with the use of mammalian becomes necessary to develop and implementation of alternative animal models. Caenorhabditis elegans is a nematode widely used as an infection model for several important human pathogens, screening for antifungal agent discovery and study of innate immunity. The limitations of the conventional methodologies concerning spatial and temporal control between host and the microorganism lead to the development of optical tweezers techniques to investigate host-pathogen interaction and the dynamic of biological process. The use of optical traps or optical tweezers is increasingly in biological research to capture and physically manipulate cells. Recently the development of a system composed by optical tweezers responsible for the movement of multiple objects and a confocal microscope for three-dimensional image acquisition permit the study of Drechmeria coniospora - C. elegans study to investigate precisely the adhesion of the fungus to C. elegans. In this context optical traps - image show a promising technology to study adhesion and elucidate the role of specific proteins as¬ 14-3-3 family using in vivo host-pathogens model. Moreover the 14-3-3 protein sequence similarity between D. coniospora and Paracoccidioides brasiliensis is 79%, permitting the use of this well-established model in this study. (AU)