Expression of Two Rye CENH3 Variants and Their Loading into Centromeres
Abstract
:1. Introduction
2. Results
2.1. Molecular Organization of the CENH3 Locus in Rye
2.2. Regulatory Regions of the CENH3 Genes
2.3. Transcription of the CENH3 Genes
2.4. Subdomain Organization of Nucleosomes Containing Different CENH3 Variants in Centromeric Chromatin
3. Discussion
4. Materials and Methods
4.1. Identification of the CENH3 Locus in the Rye Genome; Analysis of Its DNA Sequences
4.2. Identification of Functional Sites in the Vicinity of the αCENH3 and βCENH3
4.3. Plant Material
4.4. RNA Extraction and cDNA Synthesis; RT-qPCR
4.5. Synthesis of Antibodies
4.6. Slide Preparation and Indirect Immunostaining
4.7. Microscopy
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
- Van de Peer, Y.; Mizrachi, E.; Marchal, K. The evolutionary significance of polyploidy. Nat. Rev. Genet. 2017, 18, 411–424. [Google Scholar] [CrossRef]
- Flagel, L.E.; Wendel, J.F. Gene duplication and evolutionary novelty in plants. New Phytol. 2009, 183, 357–364. [Google Scholar] [CrossRef] [PubMed]
- Jiang, X.; Assis, R. Rapid functional divergence after small-scale gene duplication in grasses. BMC Evol. Biol. 2019, 19, 97. [Google Scholar] [CrossRef] [PubMed]
- Force, A.; Lynch, M.; Pickett, F.B.; Amores, A.; Yan, Y.-I.; Postlethwait, J. Preservation of duplicate genes by complementary, degenerative mutations. Genetics 1999, 151, 1531–1545. [Google Scholar] [CrossRef] [PubMed]
- Hittinger, C.T.; Carroll, S.B. Gene duplication and the adaptive evolution of a classic genetic switch. Nature 2007, 449, 677–681. [Google Scholar] [CrossRef]
- Rastogi, S.; Liberles, D.A. Subfunctionalization of duplicated genes as a transition state to neofunctionalization. BMC Evol. Biol. 2005, 5, 28. [Google Scholar] [CrossRef] [Green Version]
- Qiu, Y.; Tay, Y.V.; Ruan, Y.; Adams, K.L. Divergence of duplicated genes by repeated partitioning of splice forms and subcellular localization. New Phytol. 2020, 225, 1011–1022. [Google Scholar] [CrossRef] [Green Version]
- Zhong, C.X.; Marshall, J.B.; Topp, C.; Mroczek, R.; Kato, A.; Nagaki, K.; Birchler, J.A.; Jiang, J.M.; Dawe, K.R. Centromeric retroelements and satellites interact with maize kinetochore protein CENH3. Plant Cell 2002, 14, 2825–2836. [Google Scholar] [CrossRef]
- Nagaki, K.; Cheng, Z.K.; Yang, S.O.; Talbert, P.B.; Kim, M.; Jones, K.M.; Henikoff, S.; Buell, C.R.; Jiang, J.M. Sequencing of a rice centromere uncovers active genes. Nat Genet. 2004, 36, 138–145. [Google Scholar] [CrossRef]
- Sanei, M.; Pickering, R.; Kumke, K.; Nasuda, S.; Houben, A. Loss of centromeric histone H3 (CENH3) from centromeres precedes uniparental chromosome elimination in interspecific barley hybrids. Proc. Nat. Acad. Sci. USA 2011, 108, 498–505. [Google Scholar] [CrossRef] [Green Version]
- Yuan, J.; Guo, X.; Hu, J.; Lv, Z.; Han, F. Characterization of two CENH3 genes and their roles in wheat evolution. New Phytol. 2015, 206, 839–851. [Google Scholar] [CrossRef]
- Evtushenko, E.V.; Elisafenko, E.A.; Gatzkaya, S.S.; Lipikhina, Y.A.; Houben, A.; Vershinin, A.V. Conserved molecular structure of the centromeric histone CENH3 in Secale and its phylogenetic relationships. Sci. Rep. 2017, 7, 17628. [Google Scholar] [CrossRef]
- Maheshwari, S.; Tan, E.H.; West, A.; Franklin, F.C.; Comai, L.; Chan, S.W. Naturally occurring differences in CENH3 affect chromosome segregation in zygotic mitosis of hybrids. PLoS Genet. 2015, 11, e1004970. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Black, B.E.; Foltz, D.R.; Chakravarthy, S.; Luger, K.; Woods, V.L.; Cleveland, D.W. Structural determinants for generating centromeric chromatin. Nature 2004, 430, 578–582. [Google Scholar] [CrossRef]
- Elisafenko, E.A.; Evtushenko, E.V.; Vershinin, A.V. The Origin and Evolution of a Two-Component System of Paralogous Genes Encoding the Centromeric Histone CENH3 in Cereals. Res. Sq. 2021. Available online: https://www.researchsquare.com/article/rs-409648/v1 (accessed on 20 March 2021). [CrossRef]
- Ishii, T.; Karimi-Ashtiyani, R.; Banaei-Moghaddam, A.M.; Schubert, V.; Fuchs, J.; Houben, A. The differential loading of two barley CENH3 variants into distinct centromeric substructures is cell type- and development-specific. Chromosome Res. 2015, 23, 277–284. [Google Scholar] [CrossRef] [PubMed]
- Rabanus-Wallace, M.T.; Hackauf, B.; Mascher, M.; Lux, T.; Wicker, T.; Gundlach, H.; Baez, M.; Houben, A.; Mayer, K.F.X.; Guo, L.; et al. Chromosome-scale genome assembly provides insights into rye biology, evolution and agronomic potential. Nat. Genet. 2021, 53, 564–573. [Google Scholar] [CrossRef]
- Li, G.; Wang, L.; Yang, J.; He, H.; Jin, H.; Li, X.; Ren, T.; Ren, Z.; Li, F.; Han, X.; et al. A high-quality genome assembly highlights rye genomic characteristics and agronomically important genes. Nat. Genet. 2021, 53, 574–584. [Google Scholar] [CrossRef]
- Neumann, P.; Pavlıková, Z.; Koblıžková, A.; Fuková, I.; Jedličková, V.; Novák, P.; Macas, J. Centromeres off the hook: Massive changes in centromere size and structure following duplication of CenH3 gene in Fabeae species. Mol. Biol. Evol. 2015, 32, 1862–1879. [Google Scholar] [CrossRef] [Green Version]
- Finseth, F.R.; Dong, Y.; Saunders, A.; Fishman, L. Duplication and adaptive evolution of a key centromeric protein in Mimulus, a genus with female meiotic drive. Mol. Biol. Evol. 2015, 32, 2694–2706. [Google Scholar] [CrossRef] [Green Version]
- Kawabe, A.; Nasuda, S.; Charlesworth, D. Duplication of centromeric histone H3 (HTR12) gene in Arabidopsis halleri and A. lyrata, plant species with multiple centromeric satellite sequences. Genetics 2006, 174, 2021–2032. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Wicker, T.; Sabot, F.; Hua-Van, A.; Bennetzen, J.L.; Capy, P.; Chalhoub, B.; Flavell, A.; Leroy, P.; Morgante, M.; Panaud, O.; et al. A unified classification system for eukaryotic transposable elements. Nat. Rev. Genet. 2007, 8, 973–982. [Google Scholar] [CrossRef] [PubMed]
- Wicker, T.; Gundlach, H.; Schulman, A.H. The Repetitive Landscape of the Barley Genome. In The Barley Genome; Stein, N., Muehlbauer, J.G., Eds.; Springer: Cham, Switzerland, 2018; pp. 123–138. [Google Scholar]
- Hernandez-Garcia, C.M.; Finer, J.J. Identification and validation of promoters and cis-acting regulatory elements. Plant Sci. 2014, 217–218, 109–119. [Google Scholar]
- Haberle, V.; Stark, A. Eukaryotic core promoters and the functional basis of transcription initiation. Nat. Rev. Mol. Cell Biol. 2018, 19, 621–637. [Google Scholar] [CrossRef]
- Roy, A.L.; Singer, D.S. Core promoters in transcription: Old problem, new insights. Trends Biochem. Sci. 2015, 40, 165–171. [Google Scholar] [CrossRef] [Green Version]
- Shahmuradov, I.A.; Umarov, R.K.; Solovyev, V.V. TSSRlant: A new tool for prediction of plant Pol II promoters. Nucl. Acids Res. 2017, 45, e65. [Google Scholar]
- Eulgem, T.; Rushton, P.J.; Somssich, I.E. The WRKY superfamily of plant transcription factors. Trends Plant Sci. 2000, 5, 199–206. [Google Scholar] [CrossRef]
- Muller, F.; Tora, L. Chromatin and DNA sequences in defining promoters for transcription initiation. Biochim. Biophys. Acta 2014, 1839, 118–128. [Google Scholar] [CrossRef]
- Shen, Q.J.; Casaretto, J.A.; Zhang, P.; Ho, T.-H.D. Functional definition of ABA-response complexes: The promoter units necessary and sufficient for ABA induction of gene expression in barley (Hordeum vulgare L.). Plant Mol. Biol. 2004, 54, 111–124. [Google Scholar] [CrossRef]
- Prieto, P.; Santos, A.P.; Moore, G.; Shaw, P. Chromosomes associate premeiotically and in xylem vessel cells via their telomeres and centromeres in diploid rice (Oryza sativa). Chromosoma 2004, 112, 300–307. [Google Scholar] [CrossRef] [Green Version]
- Luger, K.; Mader, A.W.; Richmond, R.K.; Sargent, D.F.; Richmond, T.J. Crystal structure of the nucleosome core particle at 2.8 angstrom resolution. Nature 1997, 389, 251–260. [Google Scholar] [CrossRef] [PubMed]
- Vershinin, A.V.; Heslop-Harrison, J.S. Comparative analysis of the nucleosomal structure of rye, wheat and their relatives. Plant Mol. Biol. 1998, 36, 149–161. [Google Scholar] [CrossRef] [PubMed]
- Assis, R.; Bachtrog, D. Neofunctionalization of young duplicate genes in Drosophila. Proc. Natl. Acad. Sci. USA 2013, 110, 17409–17414. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Assis, R.; Bachtrog, D. Rapid divergence and diversification of mammalian duplicate gene function. BMC Evol. Biol. 2015, 15, 138. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Karimi-Ashtiyani, R.; Ishii, T.; Niessen, M.; Stein, N.; Heckmann, S.; Gurushidze, M.; Banaei-Moghaddam, A.M.; Fuchs, J.; Schubert, V.; Koch, K.; et al. Point mutation impairs centromeric CENH3 loading and induces haploid plants. Proc. Natl. Acad. Sci. USA 2015, 112, 11211–11216. [Google Scholar] [CrossRef] [Green Version]
- Kursel, L.E.; Malik, H.S. Recurrent gene duplication leads to diverse repertoires of centromeric histones in Drosophila species. Mol. Biol. Evol. 2017, 34, 1445–1462. [Google Scholar] [CrossRef] [Green Version]
- Ishii, T.; Juranić, M.; Maheshwari, S.; Bustamante, F.O.; Vogt, M.; Salinas-Gamboa, R.; Dreissig, S.; Gursanscky, N.; How, T.; Demidov, D.; et al. Unequal contribution of two paralogous CENH3 variants in cowpea centromere function. Commun. Biol. 2020, 3, 775. [Google Scholar] [CrossRef]
- Bodor, D.L.; Mata, J.F.; Sergeev, M.; David, A.F.; Sallmian, K.J.; Panchenko, T.; Cleveland, D.W.; Black, B.E.; Shah, J.V.; Jansen, L.E. The quantitative architecture of centromeric chromatin. eLife 2014, 3, e02137. [Google Scholar] [CrossRef] [Green Version]
- Shang, W.H.; Hori, T.; Martins, N.M.; Toyoda, A.; Misu, S.; Monma, N.; Hiratani, I.; Maeshima, K.; Ikeo, K.; Fujiyama, A.; et al. Chromosome engineering allows the efficient isolation of vertebrate neocentromeres. Dev. Cell 2013, 24, 635–648. [Google Scholar] [CrossRef] [Green Version]
- Neumann, P.; Schubert, V.; Fuková, I.; Manning, J.E.; Houben, A.; Macas, J. Epigenetic histone marks of extended meta-polycentric centromeres of Lathyrus and Pisum chromosomes. Front. Plant Sci. 2016, 7, 234. [Google Scholar] [CrossRef] [Green Version]
- Blower, M.D.; Sullivan, D.A.; Karpen, G.H. Conserved organization of centromeric chromatin in flies and human. Dev. Cell 2002, 2, 319–330. [Google Scholar] [CrossRef] [Green Version]
- Sullivan, B.A.; Karpen, G.H. Centromeric chromatin exhibits a histone modification pattern that is distinct from both euchromatin and heterochromatin. Nat. Struct. Mol. Biol. 2004, 11, 1076–1083. [Google Scholar] [CrossRef] [PubMed]
- Gish, W. Advanced Biocomputing 1996–2019. Available online: https://blast.advbiocomp.com (accessed on 24 August 2020).
- Smit, A.F.A.; Hubley, R.; Green, P. RepeatMasker Open-4.0. 2013–2015. Available online: http://www.repeatmasker.org (accessed on 5 February 2014).
- Kearse, M.; Moir, R.; Wilson, A.; Stones-Havas, S.; Cheung, M.; Sturrock, S.; Buxton, S.; Cooper, A.; Markowitz, S.; Duran, C.; et al. Geneious Basic: An integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics 2012, 28, 1647–1649. [Google Scholar] [CrossRef] [PubMed]
- Edgar, R.C. MUSCLE: Multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res. 2004, 32, 1792–1797. [Google Scholar] [CrossRef] [Green Version]
- Solovyev, V.V.; Shahmuradov, I.A.; Salamov, A.A. Identification of Promoter Regions and Regulatory Sites. In Computational Biology of Transcription Factor Binding; Part of the Methods in Molecular Biology Book Series; Ladunga, I., Ed.; Springer: New York, NY, USA; Dordrecht, The Netherland; Heidelberg, Germany; London, UK, 2010; Volume 674, pp. 57–83. [Google Scholar]
- Shahmuradov, I.A.; Solovyev, V.V. Nsite, NsiteH and NsiteM computer tools for studying transcription regulatory elements. Bioinformatics 2015, 31, 3544–3545. [Google Scholar] [CrossRef] [Green Version]
- Zadoks, J.C.; Chang, T.T.; Konzak, C.F. A decimal code for the growth stages of cereals. Weed Res. 1974, 14, 415–421. [Google Scholar] [CrossRef]
- Paolacci, A.R.; Tanzarella, O.A.; Porceddu, E.; Ciaffi, M. Identification and validation of reference genes for quantitative RT-PCR normalization in wheat. BMC Mol. Biol. 2009, 10, 11. [Google Scholar] [CrossRef] [Green Version]
- Larionov, A.; Krause, A.; Miller, W. A standard curve based method for relative real time PCR data processing. BMC Bioinform. 2005, 6, 62. [Google Scholar] [CrossRef] [Green Version]
- Wu, D.; Ruban, A.; Fuchs, J.; Macas, J.; Novák, P.; Vaio, M.; Zhou, Y.; Houben, A. Nondisjunction and unequal spindle organization accompany the drive of Aegilops speltoides B chromosomes. New Phytol. 2019, 223, 1340–1352. [Google Scholar] [CrossRef]
- Jasencakova, Z.; Meister, A.; Walter, J.; Turner, B.M.; Schubert, I. Histone H4 acetylation of euchromatin and heterochromatin is cell cycle dependent and correlated with replication rather than with transcription. Plant Cell 2000, 12, 2087–2100. [Google Scholar] [CrossRef] [Green Version]
- Loureiro, J.; Rodriguez, E.; Doležel, J.; Santos, C. Comparison of four nuclear isolation buffers for plant DNA flow cytometry. Ann. Bot. 2006, 98, 679–689. [Google Scholar] [CrossRef] [PubMed]
- Vaskova, E.A.; Dementyeva, E.V.; Shevchenko, A.I.; Pavlova, S.V.; Grigor’eva, E.V.; Zhelezova, A.I.; VandeBerg, J.L.; Zakian, S.M. Dynamics of the Two Heterochromatin Types during Imprinted X Chromosome Inactivation in Vole Microtus levis. PLoS ONE 2014, 9, e88256. [Google Scholar]
- Houben, A.; Schroeder-Reiter, E.; Nagaki, K.; Nasuda, S.; Wanner, G.; Murata, M.; Endo, T.R. CENH3 interacts with the centromeric retrotransposon cereba and GC-rich satellites and locates to centromeric substructures in barley. Chromosoma 2007, 116, 275–283. [Google Scholar] [CrossRef] [PubMed]
- Hesse, S.; Zelkowski, M.; Mikhailova, E.I.; Keijzer, C.J.; Houben, A.; Schubert, V. Ultrastructure and dynamics of synaptonemal complex components during meiotic pairing and synapsis of standard (A) and accessory (B) rye chromosomes. Front. Plant Sci. 2019, 10, 773. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Weisshart, K.; Fuchs, J.; Schubert, V. Structured Illumination Microscopy (SIM) and Photoactivated Localization Microscopy (PALM) to Analyze the Abundance and Distribution of RNA Polymerase II Molecules on Flow-sorted Arabidopsis Nuclei. Bio-protocol 2016, 6, e1725. [Google Scholar] [CrossRef] [Green Version]
Gypsy-Like (RLG) * | Copia-Like (RLC) * | ||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|
Family | % in IS2 | % in Superfamily (in IS2) | Family | % in Superfamily (in Genome) | % in Genome | Family | % in IS2 | % in Superfamily (in IS2) | Family | % in Superfamily (in Genome) | % in Genome |
Daniela | 22.5 | 32.3 | Sabrine | 18.2 | 8.8 | WIS | 10.4 | 44.8 | Angela | 37.38 | 5.37 |
Sabrine | 16.5 | 24.5 | Daniela | 10.3 | 4.9 | Inga | 6.2 | 26.8 | WIS | 22.65 | 3.25 |
WHAM | 9.3 | 13.8 | Erika | 6.2 | 3.0 | Angela | 5.1 | 22.1 | Barbara | 14.41 | 2.07 |
Erika | 5.3 | 7.9 | Laura | 6.0 | 2.9 | Inga | 5.15 | 0.74 | |||
Romani | 5.2 | 7.7 | Sabine | 5.8 | 2.8 | Eugene | 2.79 | 0.40 |
Types of Clusters with Variants of CENH3 | Clusters Measured | Size, nm | Size, kb * |
---|---|---|---|
Clusters containing nucleosomes with αCENH3-only signals | 97 | 278–2588 | 4.5–42.3 |
Clusters containing nucleosomes with βCENH3-only signals | 27 | 202–714 | 3.3–11.7 |
Clusters containing nucleosomes with αCENH3 and βCENH3 signals colocalized | 48 | 293–7205 | 4.8–117.9 |
Size of gaps between clusters with CENH3 signals | 72 | 206–5399 | 3.4–88.3 |
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Evtushenko, E.V.; Elisafenko, E.A.; Gatzkaya, S.S.; Schubert, V.; Houben, A.; Vershinin, A.V. Expression of Two Rye CENH3 Variants and Their Loading into Centromeres. Plants 2021, 10, 2043. https://doi.org/10.3390/plants10102043
Evtushenko EV, Elisafenko EA, Gatzkaya SS, Schubert V, Houben A, Vershinin AV. Expression of Two Rye CENH3 Variants and Their Loading into Centromeres. Plants. 2021; 10(10):2043. https://doi.org/10.3390/plants10102043
Chicago/Turabian StyleEvtushenko, Elena V., Evgeny A. Elisafenko, Sima S. Gatzkaya, Veit Schubert, Andreas Houben, and Alexander V. Vershinin. 2021. "Expression of Two Rye CENH3 Variants and Their Loading into Centromeres" Plants 10, no. 10: 2043. https://doi.org/10.3390/plants10102043