Elucidation of the functional role of A. thaliana KNL2 in epigenetic regulation of kinetochore assembly in plants
Final Report Abstract
In frame of this project we investigated the molecular mechanisms underlying the centromeric localization of Kinetochore Null 2 (KNL2) and Centromere Protein C (CENP-C) in plant cells. Our research reveals that the centromere-targeting domain coupled with DNA-binding regions are crucial for the precise localization of these proteins to the centromere, a finding that deepens our understanding of kinetochore assembly in plants and other organisms. Our key findings include: 1. Demonstration that the CENPC-k and CENPC motifs alone are not sufficient for the centromere targeting of KNL2 and CENP-C in Nicotiana benthamiana and Arabidopsis thaliana. 2. Identification of flanking DNA-binding regions near the CENPC-k and CENPC motifs, which are essential for their interaction with centromeric DNA. 3. Evidence that the DNA-binding ability of these proteins is critical for their anchoring at the centromere. This study provides novel insights into the molecular architecture of the plant centromere and opens new avenues for research into kinetochore structure and function. The implications of our findings extend beyond plant biology, offering potential strategies for manipulating chromosome segregation in biotechnological applications. Additionally, we showed that a T-DNA knockout mutant of KNL2 is an effective haploid inducer when crossed with Arabidopsis wild-type plants. We demonstrated that short-term exposure of knl2 to heat stress leads to an increase in the haploid induction efficiency from 1% to 10%. Moreover, the stress treatment regime defined for the haploid induction process with the knl2 mutant also appeared to be effective for the cenh3-4 mutant. Additionally, we showed that the introduction of a point mutation in the CENPC-k motif of KNL2 is sufficient to create a haploid inducer line.
Publications
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Immunolabeling of Nuclei/Chromosomes in Arabidopsis thaliana. Methods in Molecular Biology, 19-28. Springer US.
Ahmadli, Ulkar; Sandmann, Michael; Fuchs, Joerg & Lermontova, Inna
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Recurrent Plant-Specific Duplications of KNL2 and its Conserved Function as a Kinetochore Assembly Factor. Molecular Biology and Evolution, 39(6).
Zuo, Sheng; Yadala, Ramakrishna; Yang, Fen; Talbert, Paul; Fuchs, Joerg; Schubert, Veit; Ahmadli, Ulkar; Rutten, Twan; Pecinka, Ales; Lysak, Martin A. & Lermontova, Inna
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High temperature increases centromere-mediated genome elimination frequency and enhances haploid induction in Arabidopsis. Plant Communications, 4(3), 100507.
Ahmadli, Ulkar; Kalidass, Manikandan; Khaitova, Lucie Crhak; Fuchs, Joerg; Cuacos, Maria; Demidov, Dmitri; Zuo, Sheng; Pecinkova, Jana; Mascher, Martin; Ingouff, Mathieu; Heckmann, Stefan; Houben, Andreas; Riha, Karel & Lermontova, Inna
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Centromeric localization of KNL2 and CENP-C proteins in plants depends on their centromere-targeting domain and DNA-binding regions.
Yalagapati, Surya Prakash; Ahmadli, Ulkar; Sinha, Aditya; Kalidass, Manikandan; Dabravolski, Siarhei; Zuo, Sheng; Yadala, Ramakrishna; Rutten, Twan; Berr, Alexandre; Talbert, Paul & Lermontova, Inna
