Donata Wawrzycka

Publications of Donata Wawrzycka

• Arsenic and antimony transporters in eukaryotes.

  Authors: Ewa Maciaszczyk-Dziubinska, Donata Wawrzycka, Robert Wysocki

  International journal of molecular sciences. 01/2012; 13(3):3527-48.

  Arsenic and antimony are toxic metalloids, naturally present in the environment and all organisms have developed pathways for their detoxification. The most effective metalloid tolerance systems inArsenic and antimony are toxic metalloids, naturally present in the environment and all organisms have developed pathways for their detoxification. The most effective metalloid tolerance systems in eukaryotes include downregulation of metalloid uptake, efflux out of the cell, and complexation with phytochelatin or glutathione followed by sequestration into the vacuole. Understanding of arsenic and antimony transport system is of high importance due to the increasing usage of arsenic-based drugs in the treatment of certain types of cancer and diseases caused by protozoan parasites as well as for the development of bio- and phytoremediation strategies for metalloid polluted areas. However, in contrast to prokaryotes, the knowledge about specific transporters of arsenic and antimony and the mechanisms of metalloid transport in eukaryotes has been very limited for a long time. Here, we review the recent advances in understanding of arsenic and antimony transport pathways in eukaryotes, including a dual role of aquaglyceroporins in uptake and efflux of metalloids, elucidation of arsenic transport mechanism by the yeast Acr3 transporter and its role in arsenic hyperaccumulation in ferns, identification of vacuolar transporters of arsenic-phytochelatin complexes in plants and forms of arsenic substrates recognized by mammalian ABC transporters.

• The regulatory inputs controlling pleiotropic drug resistance and hypoxic response in yeast converge at the promoter of the aminocholesterol resistance gene RTA1.

  Authors: Anna Kołaczkowska, Myriam Manente, Marcin Kołaczkowski, Justyna Laba, Michel Ghislain, Donata Wawrzycka

  FEMS yeast research. 11/2011;

  Aminosterols possessing potent fungicidal activity are attractive alternatives to currently available antifungals. Although their precise mechanism of action is not fully understood, the effect ofAminosterols possessing potent fungicidal activity are attractive alternatives to currently available antifungals. Although their precise mechanism of action is not fully understood, the effect of 7-aminocholesterol (7-ACH) involves a partial block of Δ8-Δ7 isomerase and C-14 reductase. The function of RTA1 encoding the 7-transmembrane helix protein, cloned as the multicopy suppressor of 7-ACH toxicity in yeast, remains unclear. In this report, we show that Rta1p is localized in the plasma membrane and has a high rate of metabolic turnover, as revealed by fluorescence microscopy, cell fractionation and pulse-chase experiments. Analysis of the RTA1-lacZ reporter activity and deletion mapping of the promoter allowed the identification of the regions responsible for negative regulation by Tup1 and the two synergistically acting repressors of hypoxic genes, Rox1p and Mot3p. This was in line with increased RTA1-mediated resistance to 7-ACH under hypoxic conditions, associated with increased Rta1p level. Overexpression of RTA1 also affected the response to the signalling sphingolipid precursor phytosphingosine. Positive inputs of two transcriptional activators Pdr1p and Upc2p were also detected, indicating a regulatory link common to sterol biosynthetic genes as well as those involved in pleiotropic drug resistance and sphingolipid metabolism.

• [Yeast as a model for studying neurodegeneration].

  Authors: Donata Wawrzycka

  Postȩpy higieny i medycyny doświadczalnej (Online). 01/2011; 65:328-37.

  At the level of genetics and physiology the yeast Saccharomyces cerevisiae is are the best characterized eukaryotic cells. The yeast cells can be used as a model to study the mechanisms involved inAt the level of genetics and physiology the yeast Saccharomyces cerevisiae is are the best characterized eukaryotic cells. The yeast cells can be used as a model to study the mechanisms involved in human disease. Yeast shares conserved cellular mechanisms with all eukaryotes including mammals and human. Nowadays, despite the lack of a neural system, yeasts are successfully used in the study of neurodegenerative disorders such as Alzheimer’s disease, Huntington’s disease and Parkinson’s disease. Exquisite genetics and molecular tools used in biology allow examination of the role of yeast homologues of human genes as well as heterologous expression of human genes in yeast. Yeasts have become a suitable model to study the causes of pathological changes in protein folding, mutations and formation of aggregates.

• [Efflux-mediated antimicrobial multidrug resistance].

  Authors: Agata Jarmuła, Ewa Obłąk, Donata Wawrzycka, Jan Gutowicz

  Postȩpy higieny i medycyny doświadczalnej (Online). 01/2011; 65:216-27.

  Multidrug resistance is a major problem in the treatment of infectious diseases caused by bacteria and fungi. One of the basic mechanisms of resistance is active efflux of distinct drugs from cells.Multidrug resistance is a major problem in the treatment of infectious diseases caused by bacteria and fungi. One of the basic mechanisms of resistance is active efflux of distinct drugs from cells. Export of toxic compounds from bacterial cells is mediated by proteins of 5 distinct families: MF, SMR, ABC, RND and MATE. The substrate spectrum of efflux pumps includes antibiotics, chemotherapeutics and detergents. Genes that determine resistance can be located on chromosomes or mobile elements (plasmids, transposons, integrons). The presence of resistance genes on mobile elements enables bacteria to transfer those genes between cells and spread the multidrug resistance phenotype. There are several inhibitors of efflux pumps that are currently in the experimental phase. Proteins that mediate multidrug resistance are also present in fungal cells. They belong mainly to the ABC superfamily of transporters and PDR subfamily. These efflux pumps are widely investigated in Saccharomyces cerevisiae.

• Vmr 1p is a novel vacuolar multidrug resistance ABC transporter in Saccharomyces cerevisiae.

  Authors: Donata Wawrzycka, Iwona Sobczak, Grzegorz Bartosz, Tomasz Bocer, Stanisław Ułaszewski, André Goffeau

  FEMS yeast research. 11/2010; 10(7):828-38.

  The Saccharomyces cerevisiae Yhl035p/Vmr1p is an ABC transporter of the MRP subfamily that is conserved in all post Whole Genome Duplication species. The deletion of the YHL035 gene caused growthThe Saccharomyces cerevisiae Yhl035p/Vmr1p is an ABC transporter of the MRP subfamily that is conserved in all post Whole Genome Duplication species. The deletion of the YHL035 gene caused growth sensitivity to several amphiphilic drugs such as cycloheximide, 2,4-dichlorophenoxyacetic acid, 2,4-dinitrophenol as well as to cadmium and other toxic metals. Vmr1p-GFP was located in the vacuolar membrane. The ATP-dependent transport of a DNP-S-glutathione conjugate was reduced in a vesicular fraction from the VMR1 deletant. The energy-dependent efflux of rhodamine 6G was increased by VMR1 deletion. Growth sensitivity to cadmium of the VMR1-deleted strain was more pronounced in glycerol/ethanol than in glucose-grown cells. The VMR1 promoter had higher activity when grown in glycerol/ethanol compared with glucose. In glucose, the VMR1 promoter was activated by the deletion of the glucose-dependent repressor ADR1.

• The yeast permease Acr3p is a dual arsenite and antimonite plasma membrane transporter.

  Authors: Ewa Maciaszczyk-Dziubinska, Donata Wawrzycka, Ewa Sloma, Magdalena Migocka, Robert Wysocki

  Biochimica et biophysica acta. 11/2010; 1798(11):2170-5.

  The Acr3p permease from the yeast Saccharomyces cerevisiae is a prototype member of the arsenical resistance-3 (Acr3) family of transporters, which are found in all domains of life. Remarkably littleThe Acr3p permease from the yeast Saccharomyces cerevisiae is a prototype member of the arsenical resistance-3 (Acr3) family of transporters, which are found in all domains of life. Remarkably little is known about substrate specificity, localization and regulation of Acr3 proteins. Here, we show that the yeast Acr3p mediates not only high-level resistance to arsenite but also moderate tolerance to antimonite. The acr3 deletion mutant shows increased sensitivity to antimonite. In addition, overexpression of the ACR3 gene complements antimonite sensitivity of cells lacking the vacuolar ABC transporter Ycf1p. Moreover, both antimonite and arsenite induce transcription of the ACR3 gene resulting in the accumulation of Acr3 transporter at the plasma membrane. However, antimonite is much weaker inducer of the ACR3 gene transcription comparing to arsenite. Interestingly, the presence of metalloids does not influence either stability of Acr3 protein or its intracellular localization suggesting that Acr3p is mainly regulated at the transcriptional level. Finally, transport experiments confirmed that Acr3p indeed mediates efflux of antimonite and thus possesses a dual arsenite and antimonite specificity.

• A novel phenotype of eight spores asci in deletants of the prion-like Rnq1p in Saccharomyces cerevisiae.

  Authors: Gabriela Orlowska-Matuszewska, Donata Wawrzycka

  Biochemical and biophysical research communications. 03/2006; 340(1):190-3.

  We report that a null rnq1 mutation in the yeast RNQ1 (YCL028w) prion-like gene of so far unknown function produces the doubling of spores in the asci. This phenotype is possibly due to the lack ofWe report that a null rnq1 mutation in the yeast RNQ1 (YCL028w) prion-like gene of so far unknown function produces the doubling of spores in the asci. This phenotype is possibly due to the lack of inhibition by Rnq1p of an additional mitotic division during ascus formation. This novel phenotype termed "octopus asci" could be similar to prion [PIN+] phenotype.