CONTACTS
Institute of Microbiology of AS CR, v.v.i.
Vídeňská 1083
142 20 Prague 4
Czech Republic
PHONE: +420 241 062 503
FAX: +420 241 062 501
MEMBERS
- Jiří Hašek, PhD. (head) hasek [at] biomed.cas.cz +420 241 062 503
- Ivana Malcová-Janatová, PhD. janatova [at] biomed.cas.cz +420 241 062 769
- Miroslava Opekarová, PhD. opekaro [at] biomed.cas.cz +420 241 062 504
- Ivana Frýdlová, PhD. frydlova [at] biomed.cas.cz +420 241 062 503
- Pavla Vašicová, PhD. vasicova [at] biomed.cas.cz +420 241 062 504
- Tomáš Groušl, MSc. (PhD student) grousl [at] biomed.cas.cz +420 241 062 590
- Lenka Senohrábková, MSc. (PhD student) senohra [at] biomed.cas.cz +420 241 062 590
- Renata Slabá, BSc. (diploma student) rslaba [at] biomed.cas.cz +420 241 062 590
- Lenka Nováková, MSc. (part-time technician) lenkan [at] biomed.cas.cz +420 241 062 590
- Jarmila Serbousková (part-time technician) +420 241 062 503
- Eva Streiblová, PhD. DSc. streiblo [at] biomed.cas.cz +420 241 062 351
- Dana Janošková, MSc. (maternity leave)
RESEARCH TOPICS
Studies on the yeast
Saccharomyces cerevisiae concern (i) the plasma
membrane compartmentalization, (ii) deregulation of
a daughter-cell specific gene DSE1 and (iii) the heat stress-induced rearrangement of
translation machinery.
CURRENT PROJECTS
Plasma membrane compartmentalization
In cooperation with the Institute of Cell Biology
and Plant Physiology at Regensburg University, we obtained
important results with a key impact on understanding of plasma
membrane arrangement in eukaryotic cells. With the use of
fluorescent markers attached to various plasma membranes
proteins, we showed that the plasma membrane of S. cerevisiae
is subdivided into at least two stable lateral compartments.
One consists of 50 to 80 patches of about 300 nm in diameter is
called MCC (Membrane Compartment of Can1, the arginine
permease). The other one- MCP (Membrane Compartment of Pma1,
the H+/ATPase) fills the space in between the MCC
patches. Some other PM proteins, like Hxt1 or Gap1 are
distributed homogeneously. The distribution of proton symporters
in MCC can be affected by, e.g., lipid composition or the plasma
membrane energization, the distribution of other proteins is
resistant to these effects. Recently we have documented that the
dwelling in the MCC protects its residents against endocytosis.
The physiological importance of the compartmentalization of the
plasma membrane is under intensive study in our group.
Deregulation of a daughter-cell specific gene DSE1
Strains of Saccharomyces cerevisiae lacking
Isw2, the catalytic subunit of the Isw2 chromatin remodeling
complex, show the mating type independent activation of the cell
wall integrity (CWI) signaling pathway. Since the CWI pathway
activation usually reflects cell wall defects, we searched for
the cell wall-related genes changed in expression. The genes
DSE1, CTS1 and CHS1 were upregulated as a result
of the Isw2 absence, according to our previously published gene
expression profiles. Western blot analyses of double deletion
mutants, however, did not indicate contribution of the chitin
metabolism-related genes CTS1 and CHS1 to the CWI pathway
activation. Nevertheless, deletion of the DSE1 gene encoding a
daughter cell-specific protein with unknown function suppressed
the CWI pathway activation in isw2Δ cells. In addition,
deletion of DSE1 also abolished the budding-within-the-birth-scar
phenotype of isw2Δ cells. Plasmid-driven overexpression
proved that deregulation of the Dse1 synthesis was also responsible
for the CWI pathway activation and manifestation of the
budding-within-the-birth-scar phenotype in wild-type cells.
The overproduced Dse1-GFP localized to both sides of the septum
and persisted in unbudded cells. Although the exact cellular role
of this daughter cell-specific protein has to be elucidated,
our data point to involvement of Dse1 in bud site selection in
haploid cells.
Stress-induced rearrangement of translation machinery
In higher eukaryotic cells exposed to various environmental stresses translation initiation factors accumulate in cytoplasmic aggregates, called stress granules. The stress granules were identified in heat-stressed fission yeast but until now their formation has not been referred for budding yeast Saccharomyces cerevisiae.
We
constructed various S. cerevisiae strains expressing fusion
proteins tagged with GFP and/or mRFP from their
chromosomal sites and analyzed redistribution of translational factors,
ribosomal proteins, mRNA binding proteins and mRNA under various
stresses. Together, our data indicate that the aggregates induced by
robust heat shock represent dynamic structures composed of some
translational components and that these structures resemble stress
granules described in higher eukaryotic cells.
FUNDING
The projects are financially supported by the following institutions:
Institutional Research Concept AV0Z50200510
Czech Science Foundation: 204/09/1924
Technology Agency of the Czech Republic: TA01011467
CR Ministry of Education, Youth and Sports: LC545
COLLABORATION
Sao Paulo State University, Araraquara, Brasil (Juliana Sposto Avaca Crusca, MSc.)
University of Regensburg, Germany (Prof. W.Tanner)
University of Graz, Austria (Prof. S.D. Kohlwein)
NICHD NIH, Bethesda, USA (Dr. A. Hinnebusch)
University of Salzburg, Austria (Prof. M. Breitenbach)
Institute of Radio Engineering and Electronics AS CR, Prague (Dr. J.Pokorný)
St John's University, New York, USA (Dr.A. Vancura)
Moscow State Univ., Moscow, Russia (Dr.P.A. Ivanov)
Technion - Israel Institute of Technology, Haifa, Israel (Dr. Mordechai (Motti) Choder)
NUI Maynooth, Maynooth, Ireland (Dr. Gary Jones)
University of Manchester, Manchester, United Kingdom (Dr. Mark Ashe)
Inst. Exp. Medicine AS CR, Prague, Czech Republic, (Dr. Jan Malinsky)
PUBLICATIONS - since 2005
Tanner W., Malinsky J., Opekarova M. (2011) In plant and animal cells, detergent-resistant membranes do not define functional membrane rafts.Plant Cell 23, 1191-1193
Streiblová E., Gryndlerová H., Valda S., Gryndler M. (2010) Tuber aestivum - hypogenous fungus neglected the Czech Republic. Czech Mycology 61, 163-173
Opekarova M., Malinsky J., Tanner W. (2010) Plants and fungi in the era of heterogenous plasma membranes. Plant Biology 12, 94-98
Loibl M., Grossmann G., Stradalova V., Klingl A., Rachel R., Tanner W., Malinsky J., Opekarova M. (2010) C-terminus of Nce102 determines the structure and function of microdomains in the yeast plasma membrane.Eukaryotic Cell 9, 1184-1192
Malinsky J., Opekarova M., Tanner W. (2010) The lateral compartmentalization of the yeast plasma membrane.Yeast 8, 473-478
Novakova Z., Hubackova S., Kosar M., Janderova-Rossmeislova L., Dobrovolna J, Vasicova P., Vancurova M., Horejsi Z., Hozak P., Bartek J., Hodny Z. (2010) Cytokine expression and signaling in drug-induced cellular senescence. Oncogene. 29, 273-284
Groušl T., Ivanov P., Frýdlová I., Vašicová P., Janda F., Vojtová J., Malínská K., Malcová I., Nováková L., Janošková D., Valášek L., Hašek J. (2009) Robust heat shock induces eIF2α-phosphorylation- independent assembly of stress granules containing eIF3 and 40S ribosomal subunits in budding yeast S. cerevisiae. J.Cell.Sci.122, 2078-2088
Frýdlová I., Malcová I., Vašicová P., Hašek J. (2009) Deregulation of DSE1 gene expression results in aberrant budding within the birth scar and CWI pathway activation in Saccharomyces cerevisiae. Eukaryot.Cell 8, 586-594
Prevorovský M., Groušl T., Stanurová J., Rynes J., Nellen W., Půta F., Folk P. (2009) Cbf11 and Cbf12, the fission yeast CSL proteins, play opposing roles in cell adhesion and coordination of cell and nuclear division. Exp Cell Res. 315, 1533-1547
Jelínek F., Cifra M., Pokorný J., Vaniš J., Šimša J., Hašek J., Frýdlová I. (2009) Measurement of electrical oscillations and mechanical vibrations of yeast cells membrane around 1 kHz. Electromagn Biol Med. 28, 223-232
Grossmann G. , Malinsky J., Stahlschmidt W., Loibl M., Weig-Meckl I., Frommer W.B., Opekarová M., Tanner W. (2008) Plasma membrane microdomains regulate turnover of transport proteins in yeast J.Cell. Biol. 183, 1075-1088
Zhang F., Gaur N.A., Hasek J., Kim S-J., Qiu H., Swanson M.J., Hinnebusch A.G. (2008) Disrupting Vesicular Trafficking at the Endosome Attenuates Transcriptional Activation by Gcn4. Mol. Cell. Biol. 28, 6796-6818
Pokorny J., Hašek J., Vanis J., Jelínek F. (2008) Biophysical aspects of cancer-electromagnetic mechanism. Indian J. Exp. Biol. 46, 310-321
Dhonukshe P., Grigoriev I., Fischer R., Tominaga M., Robinson D.G., Hašek J. , Paciorek T., Petrášek J., Seifertová D., Zažímalová E., Gadella Jr T.W.J., Stierhof Y-D., Ueda T., Oiwa K., Akhmanova A., Brock R., Spang A., Friml J. (2008) Auxin transport inhibitors impair vesicle motility and actin cytoskeleton dynamics in diverse eukaryotes. PNAS 105, 4489-4494
Frýdlová I., Basler M., Vašicová P., Malcová I., Hašek J. (2007) Special type of pheromone-induced invasive growth in Saccharomyces cerevisiae. Current Genetics 52, 87-96
Grossmann G., Opekarová M., Malínský J., Weig-Meckl I.,Tanner W. (2007) Membrane potential governs lateral segregation of plasma membrane proteins and lipids in yeast. EMBO J. 26, 1-8
Grossmann G., Opekarová M., Nováková L., Stolz J., Tanner W. (2006) Lipid Raft-Based Membrane Compartmentation of a Plant Transport Protein Expressed in Saccharomyces cerevisiae. Eukaryotic Cell 5, 945-953
Malcová-Janatová I., Koubek Z., Malínská K., Raková R., Hašek J. (2006) The fission yeast ortholog of eIF3a subunit is not functional in Saccharomyces cerevisiae. Folia Microbiologica 51, 555-564
Petříčková K, Hašek J., Benada O., Petříček M. (2006) The WD-40 repeat protein PkwA of Thermomonospora curvata is associated with rapid growth and is localized in the tips of growing hyphae. FEMS Microbiol. Lett. 258, 187-193
Jelínek F., Šaroch J., Kučera O., Hašek J., Pokorný J., Jafrezic-Renault N., Ponsonnet L. (2006) Measurement of electromagnetic activity of yeast cells at 42GHz. Radioengineering 15, 1-4
Nguyen P.H., Hašek J., Kohlwein S.D., Romero C., Choi J.H., Vancura A. (2005) Interaction of Pik1p and Sjl1 proteins in membrane trafficking. FEMS Yeast Research 5, 363-371
Hašek, J. (2005) Yeast fluorescence microscopy. p85-96, In: Methods in Molecular Biology, Vol. 313 :Yeast Protocols (W.Xiao ed.) Totowa, NJ: Humana Press Inc.
Jelínek F, Pokorný J, Šaroch J, Hašek J (2005) Experimental investigation of electromagnetic activity of yeast cell at millimeter waves. Electromagnetic Biology and Medicine 24, 301-307
Opekarová M., Malínská K., Nováková L., Tanner W. (2005) Differential effect of phosphatidylethanolamine depletion on raft proteins. Further evidence for diversity of rafts in Saccharomyces cerevisiae. Biochim. Biophys. Acta 1711, 87-95
Hašek J., Peřinka L., Valášek L. (2005) Specification of the monoclonal antibody PK1 reactivity in Chinese hamster ovary cells. Folia Biologica 51, 50-51
Pokorný J., Hašek J., Jelínek F. (2005) Electromagnetic field of microtubules: Effects of transfer of mass particles and electrons. J. Biol. Phys. 31, 501-514
Pokorný J., Hašek J., Jelínek F. (2005) Endogenous Electric Field and Organization of Living Matter. Electromagnetic Biology and Medicine 24, 185-197