Laboratory of Cell Membrane Physiology
Head: Elżbieta WYROBA
Staff: Paweł Kwaśniak, Karolina Miller (student)
Molecular mechanisms regulating endocytosis and degradative pathways in different cell types undergoing gene silencing (RNAi) and stimulation. Evolutionary conservancy of the trafficking and sorting mechanisms are studied at the genomic, proteomic and cellular level. Components of the machinery involved in endosome/phagosome formation and membrane targeting in model unicellular eukaryote Paramecium octaurelia. Several genes encoding proteins indispensable in these processes were cloned in our Lab (including those first shown in unicellular eukaryotes) exhibiting a high homology to respective mammalian counterparts. Our ongoing research is focused on the functional/ structural analyses of the products of these genes with special emphasis on Rab proteins and their effectors.
• quantitative/relative real time PCR
• 2D electrophoresis
• Southern/Northern hybridization
• mass spectroscopic analysis
• Western analysis
• immunolocalization in confocal and electron microscopy
• 3D tomographic modeling
Current research activities:
• studies on evolutionary conservancy of the trafficking and sorting machinery
• revealing posttranslational modifications (PTM) as the putative mechanism of neofunctionalization of the paralogous genes
• functional analysis of Rab7 isotypes upon silencing
• identifying Rab7 effectors and cooperating proteins
• analysis of components essential in the endosomal/lysososmal pathway
• an approach with point mutagenesis and recombinant proteins to study PTM
Osińska M, Wiejak J., Wypych E, Bilski H., Bartosiewicz R, Wyroba E. (2011) Distinct expression, localization and function of two Rab7 proteins encoded by paralogous genes in a free-living model eukaryote. Acta Biochim. Polon. 58: 597-607.
Liu L., Wyroba E., Satir B.H. (2011) RNAi knockdown of parafusin inhibits the secretory pathway. Eur. J. Cell Biol. 90: 844-853.
Boncela J, Przygodzka P, Papiewska-Pajak I, Wyroba E, Osinska M, Cierniewski CS (2011) Plasminogen activator inhibitor type 1 interacts with α subunit of proteasome and modulates its activity. J. Biol. Chem. 286: 6820-6831.
Boncela J, Przygodzka P, Papiewska-Pajak I, Wyroba E, Cierniewski CS (2011) Association of Plasminogen Activator Inhibitor Type 2 (PAI-2) with Proteasome within Endothelial Cells Activated with Inflammatory Stimuli. J. Biol. Chem. 286:43164-43171.
Mackiewicz P, Wyroba E (2009) Phylogeny and evolution of Rab7 and Rab9 proteins. BMC Evolutionary Biology 9:101
Wyroba E, Surmacz L, Osinska M, Wiejak (2007) Phagosome maturation in unicellular eukaryote Paramecium: the presence of RILP, Rab7 and LAMP-2 homologues. Eur J Histochem 51:163-172
Publications list: wik.nencki.gov.pl