molecular and biochemical background of mechanisms regulating the process of endo- and exocytosis: genomic and proteomic analysis.
Components of machinery involved in endosome/phagosome formation, sorting and membrane trafficking in model unicellular eukaryote Paramecium are under study at the gene, protein and cellular level.Several genes encoding proteinsindispensable in these processes have been cloned in our Laboratory and they exhibit a high homology to the respective mammalian counterparts. Ongoing research is focused on functional/structural analysis of the products of these genes.
A wide variety of methods are used: real time PCR, RT-PCR, cloning and sequencing, 2 D protein electrophoresis, mass spectroscopic analysis, Southern/Northern hybridization, fluorescent staining, fluorimetry, immunolocalization in confocal and electron microscopy, and Western blotting.
Current research activities
functional analysis of the cloned paralogous genes encoding Rab7: real-time PCR analysis of expression during endocytic processes; pattern of intracellular localization of their protein products by confocal/electron microscopic immunocytochemistry; MALDI; silencing of these genes;
looking for Rab7 effector proteins – an evidence of expression of Rab-interacting lysosomal protein at mRNA level (sequence deposited in GenBank) as the first example of its presence in unicellular eukaryotes;
biochemical, immunocytochemical and ultrastructural studies of during internalization processes;
analysis of components cooperating with dynamin and Rab7 in endosomal/lysososmal pathway;
identification of effector domains of two other components of trafficking pathway: Rab 1 and Rab 11;
receptor-mediated endocytosis (RME) versus fluid phase uptake;
identification of proteins involved in membrane targeting and fusion in endo- and exocytosis;
studies on evolutionary conservancy of the trafficking and sorting machinery.
The Laboratory is involved in the promotion of science. The whole-day open-air presentation on “How to see DNA” was during ‘Picnic of Science'. The special sessions – open to the public – devoted to the receptor-mediated uptake of low density cholesterol (LDL) and genetic disorders of this process leading to familial hyperocholesterolemia were organized during the “Festival of Science” including EU 2006 event ‘Faces behind Science – Night with Researcher' (project 044769).
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Wyroba E., Liu L., Satir B.H. (2007) Parafusin: isoforms, number of genes and evolutionary origin. FASEB J.21: 821.2
Wiejak J., Surmacz L., Wyroba E. (2006) Cloning of two genes encoding Rab7 in Paramecium. Acta Biochim. Polon. 53: 149-56
Boncela J., Smolarczyk K., Wyroba E., Cierniewski C.S. (2006) Binding of PAI-1 to endothelial cells stimulated by thymosin beta 4 and modulation of their fibrinolytic potential. J. Biol. Chem. 281: 1066-1072
Wiejak J., Surmacz L., Wyroba E. (2004) Dynamin-association with agonist-mediated sequestration of beta-adrenergic receptor in single-cell eukaryote Paramecium. J. Exp. Biol.207: 1625-1632