Laboratory of Cell Membrane Physiology
Head: Elżbieta WYROBA
Staff: Kamil Kobyłecki (student), Karolina Miller (student)
Research profile:
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.
Methods:
• quantitative/relative real time PCR
• RT-PCR
• cloning
• 2D electrophoresis
• Southern/Northern hybridization
• mass spectroscopic analysis
• fluorometry
• 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
Selected publications:
Liu L., Wyroba E., Satir B.H. (2011) RNAi knockdown of parafusin inhibits the secretory pathway. European Journal of Cell Biology, 90: 844-853.
Mackiewicz P., Wyroba E. (2009) Phylogeny and evolution of Rab7 and Rab9 proteins. BMC Evolutionary Biology, 9: 101.
Wyroba E., Surmacz L., Osińska M., Wiejak J. (2007) Phagosome maturation in unicellular eukaryote Paramecium: the presence of RILP, Rab7 and LAMP-2 homologues. European Journal of Histochemistry, 51: 163-172.
Wiejak J., Surmacz L., Wyroba E. (2006) Cloning of two genes encoding Rab7 in Paramecium. Acta Biochimica Polonica, 53: 149-156.
Wiejak J., Surmacz L., Wyroba E. (2004) Dynamin- and clathrin-dependent endocytic pathway in unicellular eukaryote Para- mecium. Biochemistry and Cell Biology, 82: 547-558.
Publications list: wik.nencki.gov.pl
