© Leibniz-Institut (IPK)

Mission

The kinetochore is located on the centromere, where tandemly arranged centromeric repeats form a compact higher-order structure. The kinetochore is a multimeric protein complex with diverse interactions between its compounds, as illustrated by the schematic model.

The research group Kinetochore Biology (KB) is interested in studying the organisation, establishment and maintenance of the kinetochore complex in plants.

The kinetochore is a large protein complex (up to 100 proteins) that assembles at specialised chromosomal domains, the centromeres, during cell divisions. The main function of this complex is to ensure the proper segregation of chromosomes during mitosis and meiosis and, consequently, genome stability in all eukaryotic organisms.

The centromere-specific histone H3-variant CenH3 is essential for kinetochore formation and centromere function. Its deposition at centromeres depends on CenH3 assembly factors, chaperones, transcription of the centromeric repeats and the epigenetic status of centromeric chromatin. Specific manipulation of the CenH3 assembly factor KNL2 yielded double haploids in Arabidopsisthaliana (I. Lermontova, WO2017/067714). The production of double haploids enables a shortcut to achieve genome homozygosity in plant breeding because it allows to produce homogeneous lines in one rather than five or more generations as in conventional breeding.

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Projects

Our main research goals are:

1. To elucidate the mechanism of kinetochore assembly and function in plants.

We focus on:

  • Deciphering the centromere-specific histone H3-variant (cenH3) loading mechanism in plants.
  • Identification and functional characterization of novel components of the plant kinetochore complex.
  • Functional analysis of protein-protein interaction networks of kinetochore components.
  • Elucidation of the mechanism of epigenetic regulation in kinetochore assembly.

 

cenH3 loading pathway

In most eukaryotes, normal kinetochore formation depends on the centromeric variant of histone H3 cenH3. The cenH3 deposition pathway in A. thaliana is represented by a three-step process: initiation, deposition and maintenance. (1) Initiation of cenH3 incorporation involves the licensing factor Kinetochore Null 2 (KNL2) and other components (Lermontova et al., 2013), (2) then cenH3 is deposited to centromeres by chaperone proteins such as NASPSIM3 (Le Goff et al., 2020).  (3) The specific mechanisms/factors for maintaining newly incorporated cenH3 are not yet known.

2.  To establish an efficient haploid induction approach based on the manipulation of kinetochore proteins.

We focus on:

  • Optimization of KNL2-based haploid induction approach in Arabidopsis and determination of its mechanism.
  • Transferring the KNL2-based haploid induction approach to crop species.

 

Generation of haploids and double haploids based on KNL2 manipulations

Crossing of A. thaliana mutant for the kinetochore protein KNL2 with wild-type results in the generation of haploids. (Lermontova, I.: Generation of haploid plants based on KNL 2. (IPK-Anmeldung), Veröffentlichung: 27.04.2017, IPK-Nr. 2015/01. WO2017/067714 (2017)).

3. To analyse the effect of environmental factors on the kinetochore assembly.

We focus on:

  • Studying the effects of environmental conditions on heterochromatin structure, kinetochore assembly and haploid induction efficiency.

 

Application of temperature and light stress affect the structure of heterochromatin

Stress conditions such as high temperature and light displayed dispersed heterochromatin structure in A. thaliana.

We apply a combination of molecular biological, biochemical, physiological, cytological, and bioinformatics tools and techniques to achieve our research goals.

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Staff

NameEmailTelephone
Scientists
Ahmadli Ulkar ahmadli@ipk-gatersleben.de+49 39482 5-511
DemidovDr. Dmitri demidov@ipk-gatersleben.de+49 39482 5-733
ORCID iD icon0000-0003-1814-1023
Kalidass Manikandan kalidass@ipk-gatersleben.de+49 39482 5-298
LermontovaDr. Inna lermonto@ipk-gatersleben.de+49 39482 5-570
ORCID iD icon0000-0003-3386-2590
Sinha Aditya sinha@ipk-gatersleben.de+49 39482 5-511
Yadala Ramakrishna yadala@ipk-gatersleben.de+49 39482 5-827
Lab Technicians
Heber Anette heber@ipk-gatersleben.de+49 39482 5-296
Kuhlmann Heike kuhlmannh@ipk-gatersleben.de+49 39482 5-583

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Publications

AuthorTitle
2021

Ahmadli U, Sandmann M, Fuchs J, Lermontova I:

Immunolabeling of nuclei/chromosomes in Arabidopsis thaliana. In: Caillaud M-C (Ed.): Plant cell division: methods and protocols, 2nd. ed. (Series: Methods in molecular biology, Vol. 2382) New York [u.a.]: Springer US (2021) in press. dx.doi.org/10.1007/978-1-0716-1744-1 ISBN 978-1-0716-1743-4

Capitao C, Tanasa S, Fulnecek J, Raxwal V K, Akimcheva S, Bulankova P, Mikulkova P, Khaitova L C, Kalidass M, Lermontova I, Scheid O M, Riha K:

A CENH3 mutation promotes meiotic exit and restores fertility in SMG7-deficient Arabidopsis. PLoS Genet. 17 (2021) e1009779. https://dx.doi.org/10.1371/journal.pgen.1009779

Houben A, Demidov D, Lermontova I, Fuchs J, Weiss O, Meier K:

Generation of haploids based on mutation of SAD2. (Patent), Veröffentlichung: 24.03.2021, IPK-Nr. 2018/06. EP3794939. (2021).

Municio C, Antosz W, Grasser K, Kornobis E, van Bel M, Eguinoa I, Coppens F, Bräutigam A, Lermontova I, Bruckmann A, Zelkowska K, Houben A, Schubert V:

The Arabidopsis condensin CAP-D subunits arrange interphase chromatin. New Phytol. 230 (2021) 972-987. https://dx.doi.org/10.1111/nph.17221

Sorge E, Demidov D, Lermontova I, Houben A, Conrad U:

Engineered degradation of EYFP-tagged CENH3 via the 26S proteasome pathway in plants. PLoS One 16 (2021) e0247015. https://dx.doi.org/10.1371/journal.pone.0247015

2020

Dvořák Tomaštíková E, Rutten T, Dvořák P, Tugai A, Ptošková K, Petrovská B, van Damme D, Houben A, Doležel J, Demidov D:

Functional divergence of microtubule-associated TPX2 family members in Arabidopsis thaliana. Int. J. Mol. Sci. 21 (2020) 2183. https://dx.doi.org/10.3390/ijms21062183

Ishii T, Juranić M, Maheshwari S, Bustamante F O, Vogt M, Salinas-Gamboa R, Dreissig S, Gursanscky N, How T, Demidov D, Fuchs J, Schubert V, Spriggs A, Vielle-Calzada J P, Comai L, Koltunow A M G, Houben A:

Unequal contribution of two paralogous CENH3 variants in cowpea centromere function. Commun. Biol. 3 (2020) 775. https://dx.doi.org://10.1038/s42003-020-01507-x

Le Goff S, Keçeli B N, Jeřábková H, Heckmann S, Rutten T, Cotterell S, Schubert V, Roitinger E, Mechtler K, Franklin F C H, Tatout C, Houben A, Geelen D, Probst A V, Lermontova I:

The H3 histone chaperone NASPSIM3 escorts CenH3 in Arabidopsis. Plant J. 101 (2020) 71-86. https://dx.doi.org/10.1111/tpj.14518

2019

Boudichevskaia A, Houben A, Fiebig A, Prochazkova K, Pecinka A, Lermontova I:

Depletion of KNL2 results in altered expression of genes involved in regulation of the cell cycle, transcription, and development in Arabidopsis. Int. J. Mol. Sci. 20 (2019) 5726. https://dx.doi.org/10.3390/ijms20225726

Demidov D, Heckmann S, Weiss O, Rutten T, Tomaštíková E D, Kuhlmann M, Scholl P, Municio C M, Lermontova I, Houben A:

Deregulated phosphorylation of CENH3 at Ser65 affects the development of floral meristems in Arabidopsis thaliana. Front. Plant Sci. 10 (2019) 928. https://dx.doi.org/10.3389/Fpls.2019.00928

Kalinowska K, Chamas S, Unkel K, Demidov D, Lermontova I, Dresselhaus T, Kumlehn J, Dunemann F, Houben A:

State-of-the-art and novel developments of in vivo haploid technologies. Theor. Appl. Genet. 132 (2019) 593–605. https://dx.doi.org/10.1007/s00122-018-3261-9

Zelkowski M, Zelkowska K, Conrad U, Hesse S, Lermontova I, Marzec M, Meister A, Houben A, Schubert V:

Arabidopsis NSE4 proteins act in somatic nuclei and meiosis to ensure plant viability and fertility. Front. Plant Sci. 10 (2019) 774. https://dx.doi.org/10.3389/fpls.2019.00774

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