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Advanced Nano and Microtechnologies Advanced Materials Structural Biology Gen. and Prot. of Plant Systems Molecular Medicine Brain and Mind Research Molecular Vet. Medicine

Functional Genomics and Proteomics of Plants - Jan Hejátko

Research Group Leader
Researcher ID
Phone: +420 54949 4165
E-mail: ,
Office:

Research areas

  • Role of cytokinins in the vascular tissue and root apical meristem formation and developmentczech bio logo nove
  • Interaction of cytokinin and other hormones, particularly auxin
  • Interaction of cytokinins with light
  • Structural basis of multistep phosphorelay signaling in plants
  • Plant proteome and phosphoproteome response to cytokinins
  • Employing the knowledge of molecular mechanisms of multistep phosphorelay-based signalling in molecular breeding
  • Production of bioactive compounds in plant production systems

Main objectives

Determination of molecular mechanisms governing hormonal regulations and their functions in plant development.

Content of research

The research group is interested in the hormonal regulation of plant development and underlying molecular mechanisms with particular emphasis on the understanding of cytokinin signalling, action and interaction with other plant growth regulators.

Plant cells are well known for their tremendous developmental plasticity. Plant hormones, particularly auxins and cytokinins were found to be major regulators of intrinsic developmental programs associated with changes of differentiation status of plant cells and tissues. That allows de novo formation of entire plants from virtually all types of specialised plant tissues. Identification of basic molecular principles involved in the regulation of plant cell division and differentiation will provide developmental model useful in the comparative biology approaches and identification of corresponding regulatory and developmental events in animal and particularly human cell systems.

In the group of Functional Genomics and Proteomics of Plants, we are interested in the molecular mechanisms underlying the regulation of plant development by plant hormones cytokinins (CKs) and their interaction with other plant hormones, e.g. auxin. In our studies we employ comprehensive approaches including forward and reverse genetics, proteomics, protein biochemistry, protein structure analysis and bioinformatics to recognise the principles of complex molecular events involved in the cytokinin signal transduction and action.

We are particularly interested in study of following problems:

  1. Interaction of auxin and cytokinins in the processes of de novo organogenesis, regulation of root meristem patterning and vascular tissue development. We are interested in the identification of molecular targets acting downstream the CK signalling pathways and the role of gene regulatory networks constituting CKdependent developmental circuits.
  2. Elucidating molecular determinants of specifi city in multistep phosphorelay (MSP), with special emphasis on the role of MSP in CK signalling. Particularly we are interested in the structural analysis of intracellular receiver domains of sensory histidine kinases that, as we have found, specifically interact with downstream signaling His-containing phosphotransfer proteins, determining thus specificity in plant MSP pathways.
  3. Hormonal regulations of plant proteome, particularly the study of CK-dependent proteome changes with the aim of identifying novel regulatory targets of CK-mediated regulations of plant development and elucidating molecular mechanisms underlying interaction of CK with other plant growth regulators, e.g. ethylene.
  4. Development and application of novel approaches, e.g. immunomodulation (production of scFv fragments recognising both cytokinins and CK signalling proteins in vivo) to modulate CK-dependent regulation of plant development.

Extending our fundamental research and use of its results in applied science and development of novel strategies useful in e.g. molecular breeding or using of plant systems for biotherapeutics production.

list / cards

Name and position

E-mail

Phone

Jan Hejátko, Ph.D.
Research Group Leader
+420 54949 4165
Blanka Pekárová, Ph.D.
Researcher
+420 54949 3029
Markéta Žďárská, Ph.D.
Postdoctoral Fellow
+420 54949 3553
Mónika Hrtyan
Laboraroty technician
Romana Hejátková
Laboratory technician
Šárka Michlíčková
Laboratory technician
+420 54949 5410
Martina Válková
Laboratory technician
+420 54949 7904
Ivana Urbánková
Specialist
Ivan Kashkan
odborný pracovník - PhD student
Jakub Zeman
Research Project Manager
+420 54949 6795
Zuzana Gelová
PhD student
Agnieszka Szmitkowska
odborná pracovnice - PhD studentka
Tomáš Faltus
technik
Amel Yamoune
odborná pracovnice - PhD studentka
Elena Zemlyanskaya
Specializovaná pracovnice
Kamil Růžička
Researcher
Aswathy Jayasree, Ph.D.
odborná pracovnice ve výzkumu - postdoc
Veronika Balakhonova
Pavel Veverka
PhD student
Kseniya Timofeyenko
odborná pracovnice - PhD studentka
Abigail Rubiato Cuyacot
Tomáš Konečný
Michal Ondruš
Petra Zvolánková
Anna Alaxinová
PhD student
Anna Bílková
Siarhei Dabravolski
PhD student
Petra Elblová
Eliška Špačková
Markéta Šámalová
senior researcher
+420 54949 8476
Gabriela Jalová
Managing assistant
+420 54949 1454

SELECTED PUBLICATIONS

2017

  • DOBISOVA, T; HRDINOVA, V; CUESTA, C; MICHLICKOVA, S; URBANKOVA, I; HEJATKOVA, R; ZADNIKOVA, P; PERNISOVA, M; BENKOVA, E; HEJATKO, J, 2017:Light Controls Cytokinin Signaling via Transcriptional Regulation of Constitutively Active Sensor Histidine Kinase CKI1. PLANT PHYSIOLOGY 174 (1), p. 387 - 404.
  • RUZICKA, K; HEJATKO, J, 2017:Auxin transport and conjugation caught together. JOURNAL OF EXPERIMENTAL BOTANY 68 (16), p. 4409 - 4412.

2016

  • NODZYNSKI, T; VANNESTE, S; ZWIEWKA, M; PERNISOVA, M; HEJATKO, J; FRIML, J, 2016:Enquiry into the Topology of Plasma Membrane-Localized PIN Auxin Transport Components. MOLECULAR PLANT 9 (11), p. 1504 - 1519.
  • PEKAROVA, B; SZMITKOWSKA, A; DOPITOVA, R; DEGTJARIK, O; ZIDEK, L; HEJATKO, J, 2016:Structural Aspects of Multistep Phosphorelay-Mediated Signaling in Plants. MOLECULAR PLANT 9 (1), p. 71 - 85.
  • PERNISOVA, M; PRAT, T; GRONES, P; HARUSTIAKOVA, D; MATONOHOVA, M; SPICHAL, L; NODZYNSKI, T; FRIML, J; HEJATKO, J, 2016:Cytokinins influence root gravitropism via differential regulation of auxin transporter expression and localization in Arabidopsis. NEW PHYTOLOGIST 212 (2), p. 497 - 509.

2015

  • DIDI, V; JACKSON, P; HEJATKO, J, 2015:Hormonal regulation of secondary cell wall formation. JOURNAL OF EXPERIMENTAL BOTANY 66 (16), p. 5015 - 5027.
  • JUPA, R; DIDI, V; HEJATKO, J; GLOSER, V, 2015:An improved method for the visualization of conductive vessels in Arabidopsis thaliana inflorescence stems. FRONTIERS IN PLANT SCIENCE 6
  • KREJCI, J; STIXOVA, L; PAGACOVA, E; LEGARTOVA, S; KOZUBEK, S; LOCHMANOVA, G; ZDRAHAL, Z; SEHNALOVA, P; DABRAVOLSKI, S; HEJATKO, J; BARTOVA, E, 2015:Post-Translational Modifications of Histones in Human Sperm. JOURNAL OF CELLULAR BIOCHEMISTRY 116 (10), p. 2195 - 2209.
  • KUDEROVA, A; GALLOVA, L; KURICOVA, K; NEJEDLA, E; CURDOVA, A; MICENKOVA, L; PLIHAL, O; SMAJS, D; SPICHAL, L; HEJATKO, J, 2015:Identification of AHK2-and AHK3-like cytokinin receptors in Brassica napus reveals two subfamilies of AHK2 orthologues. JOURNAL OF EXPERIMENTAL BOTANY 66 (1), p. 339 - 353.
  • RUZICKA, K; URSACHE, R; HEJATKO, J; HELARIUTTA, Y, 2015:Xylem development - from the cradle to the grave. NEW PHYTOLOGIST 207 (3), p. 519 - 535.
  • ZDARSKA, M; DOBISOVA, T; GELOVA, Z; PERNISOVA, M; DABRAVOLSKI, S; HEJATKO, J, 2015:Illuminating light, cytokinin, and ethylene signalling crosstalk in plant development. JOURNAL OF EXPERIMENTAL BOTANY 66 (16), p. 4913 - 4931.

2014

  • BORKOVCOVA, P; PEKAROVA, B; VALKOVA, M; DOPITOVA, R; BRZOBOHATY, B; JANDA, L; HEJATKO, J, 2014:Antibodies against CKI1(RD), a receiver domain of the sensor histidine kinase in Arabidopsis thaliana: From antigen preparation to in planta immunolocalization. PHYTOCHEMISTRY 100 , p. 6 - 15.
  • ZABRADY, M; HRDINOVA, V; MULLER, B; CONRAD, U; HEJATKO, J; JANDA, L, 2014:Targeted In Vivo Inhibition of Specific Protein-Protein Interactions Using Recombinant Antibodies. PLOS ONE 9 (10)

2013

  • BENITEZ, M; HEJATKO, J, 2013:Dynamics of Cell-Fate Determination and Patterning in the Vascular Bundles of Arabidopsis thaliana. PLOS ONE 8 (5)
  • NOVACEK, J; JANDA, L; DOPITOVA, R; ZIDEK, L; SKLENAR, V, 2013:Efficient protocol for backbone and side-chain assignments of large, intrinsically disordered proteins: transient secondary structure analysis of 49.2 kDa microtubule associated protein 2c. JOURNAL OF BIOMOLECULAR NMR 56 (4), p. 291 - 301.
  • ZD'ARSKA, M; ZATLOUKALOVA, P; BENITEZ, M; SEDO, O; POTESIL, D; NOVAK, O; SVACINOVA, J; PESEK, B; MALBECK, J; VASICKOVA, J; ZDRAHAL, Z; HEJATKO, J, 2013:Proteome Analysis in Arabidopsis Reveals Shoot- and Root-Specific Targets of Cytokinin Action and Differential Regulation of Hormonal Homeostasis. PLANT PHYSIOLOGY 161 (2), p. 918 - 930.

2012

  • DOKLADAL, L; OBORIL, M; STEJSKAL, K; ZDRAHAL, Z; PTACKOVA, N; CHALOUPKOVA, R; DAMBORSKY, J; KASPAROVSKY, T; JEANDROZ, S; ZD'ARSKA, M; LOCHMAN, J, 2012:Physiological and proteomic approaches to evaluate the role of sterol binding in elicitin-induced resistance. JOURNAL OF EXPERIMENTAL BOTANY 63 (5), p. 2203 - 2215.

2011

  • PEKAROVA, B; KLUMPLER, T; TRISKOVA, O; HORAK, J; JANSEN, S; DOPITOVA, R; BORKOVCOVA, P; PAPOUSKOVA, V; NEJEDLA, E; SKLENAR, V; MAREK, J; ZIDEK, L; HEJATKO, J; JANDA, L, 2011:Structure and binding specificity of the receiver domain of sensor histidine kinase CKI1 from Arabidopsis thaliana. PLANT JOURNAL 67 (5), p. 827 - 839.

before 2011

  • HEJATKO, J; RYU, H; KIM, GT; DOBESOVA, R; CHOI, S; CHOI, SM; SOUCEK, P; HORAK, J; PEKAROVA, B; PALME, K; BRZOBOHATY, B; HWANG, I, 2009: The histidine kinases CYTOKININ-INDEPENDENT1 and ARABIDOPSIS HISTIDINE KINASE2 and 3 regulate vascular tissue development in Arabidopsis shoots. PLANT CELL 21 (7), p. 2008 - 2021.
  • PERNISOVA, M; KLIMA, P; HORAK, J; VALKOVA, M; MALBECK, J; SPUCEK, P; REICHMAN, P; HOYEROVA, K; DUBOVA, J; FRIML, J; ZAZIMALOVA, E; HEJATKO, J, 2009: Cytokinins modulate auxin-induced organogenesis in plants via regulation of the auxin efflux. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE U.S.A. 106, p. 3609 - 3614.
  • HEJATKO, J; BLILOU, I; BREWER, PB; FRIML, J; SCHERES, B; BENKOVA, E, 2006: In situ hybridisation technique for mRNA detection in whole mount Arabidopsis samples. NATURE PROTOCOLS 1, p. 1939 - 1946.
  • HEJATKO, J; PERNISOVA, M; ENEVA, T; PALME, K; Brzobohatý, B, 2003: The putative sensor histidine kinase CKI1 is involved in female gametophyte development in Arabidopsis. MOLECULAR GENETICS AND GENOMICS 269, p. 443 - 453.


PATENTS

  • HEJATKO, J; DOBESOVA, R; DUBOVA. J; HWANG, I; RYU, H, 2009: Method of regulation of biomass production in plants. Czech patent No. 300145, Granted on January 15, 2009.  Applicants: Masaryk University and POSTECH Academy Industry Foundation

GRANTY

  • Deciphering Molecular Mechanisms Integrating Light and Hormonal Control Over Plant Development (GA15-22000S), Czech Science Foundation - Standard Grants, 2015 - 2017
  • Structural basis for the specificity of signal transduction in plants: interaction network of histidine kinase receiver domains in Arabidopsis (GAP305/11/0756), Czech Science Foundation - Standard Grants, 2011 - 2014
  • The role of cytokinins and auxin interactions in the regulation of root gravitropism in Arabidopsis thaliana (GAP501/11/1150), Czech Science Foundation - Standard Grants, 2011 - 2013
  • Immunomodulation as a functional proteomics tool for cytokinin signaling study in Arabidopsis thaliana (GA521/09/1699), Czech Science Foundation - Standard Grants, 2009 - 2012
  • Identification of molecular components and the mechanism of polar targeting of PIN auxin transport proteins in Arabidopsis thaliana (IAA601630703), Academy of Sciences of the Czech Republic - Grants of distinctly investigative character focused on the sphere of research pursued at present particularly in the Academy of Sciences of the Czech Rep., 2007 - 2011
  • Regulation of morphogenesis of plant cells and organs (LC06034), MEYS - Basic Research Centres, 2006 - 2011

CURRENT RESEARCH INFRASTRUCTURE

Equipment for cultivation of plants under tightly regulated conditions (fytotrons) and under defined light quality and quantity. Expertise and high-end instrumentation in advanced microscopy techniques that include high-end confocal microscope equipped with white laser, highly sensitive hybrid detectors and module for fluorescence life-time imaging (FLIM) and fluorescence correlation spectroscopy (FCS). Image analysis software allowing highly sensitive and specific fluorescence imaging in living cells via fluorescence intensity decay shape analysis microscopy (FIDSAM). High-throughput system for automated microscopy (Olympus “.slide”) and horizontal confocal macroscope (modified Nikon AZ-C1 system), allowing detailed, real-time fluorescent protein localisation and dynamics in vivo.

Video of High-throughput system for automated microscopy (Olympus “.slide”) can be found HERE.

supplement-figure-1

1. DIRIGENT Genes in the Control of Plant Development

Supervisor: doc. RNDr. Jan Hejátko, Ph.D.

Annotation:

In our lab we are interested in the study of plant hormonal signaling pathways, particularly cytokinins (CKs) in the development of model plant Arabidopsis thaliana (1-4). CK signaling is mediated via what is called multistep phosphorelay signaling (MSP). Recently we have found that CKs control xylem differentiation via regulation of the composition of the cell wall and its biomechanical properties (Didi et al., unpublished). Our bioinformatics as well as experimental evidence suggest that there are more direct targets of CK signaling pathway with a potential role in the control of the cell wall properties. Among these are members of the almost completely unexplored Arabidopsis thaliana DIRIGENT (AtDIR) gene family. Based on our preliminary results it seems that AtDIRs could have an important role in the plant response to both biotic and abiotic stress as well as in the control of plant organogenesis. The aim of the project is to analyze the role of AtDIR13 and AtDIR14 in the Arabidopsis development. The work will include using up-to-date genetics, molecular biology and advanced imaging approaches including the recently introduced Brillouin imaging developed for the non-invasive measurement of biomechanical cell wall properties (5). The project promises to uncover novel mechanisms underlying hormonal control of plant development of fundamental importance. We offer: Experimental work on own project with the possibility of publication in distinguished international journals, nice lab staff, world-class lab equipment, attractive environment of novel university campus, help with both intellectual and practical problems, competitive salary corresponding to the work efficiency, collaboration with abroad lab. The position will be available from January 1 2018. Contact and further info: +420 5 4949 4165, hejatko@sci.muni.cz

References: 

1. Dobisova, T., et al. (2017). Plant Physiol 174, 387-404.

2. Paniagua, C., et al. (2017). J Exp Bot.

3. Pernisova, M., et al. (2016). New Phytol 212, 497-509.

4. Zd'arska, M., et al. (2013). Plant Physiology 161, 918-930.

5. Elsayad, K., et al. (2016). Sci Signal 9, rs5.

Keywords: cytokinins, CKs, multistep phosphorelay signaling, MSP, AtDIR, Brillouin imaging, plant hormonal signaling pathways, mdel plant Arabidopsis thaliana

2. Genetic variability in multistep phoshorelay signaling in plants

Supervisor: doc. RNDr. Jan Hejátko, Ph.D.

Annotation

In our lab we are interested in the study of plant hormonal signaling pathways, particularly cytokinins in the development of model plant Arabidopsis thaliana (1-3). Cytokinin signaling is mediated via what is called multistep phosphorelay signaling (MSP). Cytokinins were described as important regulators of biotic and abiotic stresses, making members of the MSP signaling pathway attractive targets in breeding programs aimed to the improvement of crop adaptation responses. Previously we have identified sensor histidine kinases working as cytokinin receptors in oil-seed rape (4). However, our knowledge on the mechanism of action of MSP signaling in crops is very limited. In frame of the project, we will study the genetic variability in individual members of the MSP signaling pathway in Arabidopsis and oil-seed rape. The role of identified variants in the cytokinin responsiveness will be tested using transgenic plants carrying the transcriptional MSP reporters as well as transient expression in plant protoplasts. Molecular mechanisms of identified polymorphisms in cytokinin responsiveness will be studied by experimental as well as structural modeling approaches. The work will include using up-to-date genetics, molecular biology and advanced imaging approaches. The project will be solved in a tight collaboration with Photon Systems Instruments company, employing their cutting-edge plant phenotyping platform. Results with impact on both fundamental research of MSP signaling as well as high application potential are expected. We offer: Experimental work on own project with the possibility of publication in distinguished international journals, nice lab staff, world-class lab equipment, attractive environment of novel university campus, help with both intellectual and practical problems, competitive salary corresponding to the work efficiency, collaboration with abroad lab. The position will be available from Aug/Sep 2018. We expect: Highly motivated candidates with ambition of reaching top-ranked results.

References: 

1. Dobisova, T., et al. (2017). Plant Physiol 174, 387-404.

2. Pernisova, M., et al. (2016). New Phytol 212, 497-509.

3. Pernisova, M., et al. (2009). Proc Natl Acad Sci U S A 106, 3609-3614.

4. Kuderova, A., et al. (2015). Journal of Experimental Botany 66, 339-353.

Keywords: cytokinins, multistep phosphorelay signaling, MSP, cytokinin receptors, plant phenotyping,  biotic and abiotic stresses

3. Structural and functional analysis of cytokinin-regulated DIRIGENT proteins from Arabidopsis thaliana

Supervisor: Ing. Blanka Pekárová, Ph.D.
Consultant: doc. RNDr. Jan Hejátko, Ph.D.

Annotation

Our Research Group Functional Genomics and Proteomics of Plants has been investigating plant hormonal signaling pathways, particularly cytokinins and ethylene in the development of model plant Arabidopsis thaliana (1-4) for a long time. Recently, we have turned our attention to understand the molecular mechanisms mediating cytokinin regulations of the cell wall composition and thus its biomechanical properties. We found that two out of 26 Arabidopsis DIRIGENT proteins (AtDIR13 and AtDIR14) are up-regulated by cytokinins in the Arabidopsis roots and they also seem to be direct targets of cytokinin signaling (2). AtDIR6 is the only member of the family with known structure (5). DIRIGENT proteins are glycoproteins mediating biosynthesis of lignan and lignin, phenolic compounds important in plant development. Some DIRIGENT proteins were shown to be involved in the biotic and abiotic stress responses. Additionally, lignin that represents approx. 15-30% dry weight of lignocellulose has potential as a renewable source of high-value aromatic chemicals. Lignans can be used as drugs in conventional medicine. The aim of this work is to clone, express, purify and crystallize AtDIR13/14 proteins and use them in the series of in vitro enzymatic assays in order to identify their substrate specificity and putative role in the catalysis of lignin and/or lignan formation. We expect: Highly motivated candidates with ambition of reaching top-ranked results. We offer: Experimental work on own project with the possibility of publication in distinguished international journals, nice lab staff, world-class lab equipment, attractive environment of novel university campus, help with both intellectual and practical problems, competitive salary corresponding to the work efficiency. The position will be available from January 2018. Contact and further info: +420 5 4949 3029, pekarova@sci.muni.cz, hejatko@sci.muni.cz

References:

1. Dobisova, T., et al. (2017). Plant Physiol. 174, 387-404.

2. Paniagua, C., et al. (2017). J Exp Bot. 68, 3287-3301.

3. Pernisova, M., et al. (2016). New Phytol. 212, 497-509.

4. Zdarska, M., et al. (2013). Plant Physiol. 161, 918-930.

5. Gasper, R., et al. (2016). Plant Physiol. 172, 2165-2175.

Keywords: cytokinin regulations, Arabidopsis DIRIGENT proteins,AtDIR13, AtDIR14, plant model Arabidopsis thaliana, hormonal signaling pathways

4.The role of cytokinin-inducible EXPANSINs in the control of cell wall properties and development in Arabidopsis

Supervisor: Mgr. Markéta Šámalová, Ph.D.
Consultant: doc. RNDr. Jan Hejátko, Ph.D.

Annotation

In our lab we are interested in the study of plant hormonal signaling pathways, particularly cytokinins (CKs) in the development of model plant Arabidopsis thaliana (1-4). Our recent preliminary data suggest an important role for CK-regulated genes, namely EXPANSINs, in the control of cell wall (CW) properties. Besides CW critical developmental importance in plants, CW-based biomaterials are exploited in diverse human activities and have high application potential as a source of sustainable and renewable energy for the future. The aim of the project is to investigate a novel role for CKs in the regulation of CW composition and structure during CK-controlled cell differentiation. We hypothesize that CKs control cell differentiation via regulation of biomechanical properties of the CW. As an important part of the project we will employ the CRISPR-Cas9 gene-editing technology and brand-new optical method, Brillouin imaging, developed for the non-invasive measurement of biomechanical CW properties (5). We expect: Highly motivated, ambitious candidates with interest in molecular biology and plants. We offer: Experimental work on own project with the possibility of publication in top-ranked international journals, friendly lab staff, world-class equipment, attractive environment of novel university campus, help with both intellectual and practical problems, competitive salary corresponding to the work efficiency. The position is available from January 2018. Contact and further info: +420 5 4949 8476, marketa.samalova@ceitec.muni.cz hejatko@sci.muni.cz

References:

1. Dobisova, T., et al. (2017). Plant Physiol 174, 387-404.

2. Paniagua, C., et al. (2017). J Exp Bot.

3. Pernisova, M., et al. (2016). New Phytol 212, 497-509.

4. Zd'arska, M., et al. (2013). Plant Physiology 161, 918-930.

5. Elsayad, K., et al. (2016). Sci Signal 9, rs5.

Keywords: CK-regulated genes, EXPANSINs, cell wall properties control, CRISPR-Cas9 gene-editing, Brillouin imaging

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