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Article: The Emerging Role of the Cytoskeleton in Chromosome Dynamics.

Spichal, Maya / Fabre, Emmanuelle

2017 Volume 8, Page(s) 60

Abstract: Chromosomes underlie a dynamic organization that fulfills functional roles in processes like transcription, DNA repair, nuclear envelope stability, and cell division. Chromosome dynamics depend on chromosome structure and cannot freely diffuse. ... ...

Abstract	Chromosomes underlie a dynamic organization that fulfills functional roles in processes like transcription, DNA repair, nuclear envelope stability, and cell division. Chromosome dynamics depend on chromosome structure and cannot freely diffuse. Furthermore, chromosomes interact closely with their surrounding nuclear environment, which further constrains chromosome dynamics. Recently, several studies enlighten that cytoskeletal proteins regulate dynamic chromosome organization. Cytoskeletal polymers that include actin filaments, microtubules and intermediate filaments can connect to the nuclear envelope via Linker of the Nucleoskeleton and Cytoskeleton (LINC) complexes and transfer forces onto chromosomes inside the nucleus. Monomers of these cytoplasmic polymers and related proteins can also enter the nucleus and play different roles in the interior of the nucleus than they do in the cytoplasm. Nuclear cytoskeletal proteins can act as chromatin remodelers alone or in complexes with other nuclear proteins. They can also act as transcription factors. Many of these mechanisms have been conserved during evolution, indicating that the cytoskeletal regulation of chromosome dynamics is an essential process. In this review, we discuss the different influences of cytoskeletal proteins on chromosome dynamics by focusing on the well-studied model organism budding yeast.
Language	English
Publishing date	2017-05-19
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2606823-0
ISSN	1664-8021
ISSN	1664-8021
DOI	10.3389/fgene.2017.00060
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Article ; Online: Germ granule dysfunction is a hallmark and mirror of Piwi mutant sterility.

Spichal, Maya / Heestand, Bree / Billmyre, Katherine Kretovich / Frenk, Stephen / Mello, Craig C / Ahmed, Shawn

Nature communications

2021 Volume 12, Issue 1, Page(s) 1420

Abstract: In several species, Piwi/piRNA genome silencing defects cause immediate sterility that correlates with transposon expression and transposon-induced genomic instability. In C. elegans, mutations in the Piwi-related gene (prg-1) and other piRNA deficient ... ...

Abstract	In several species, Piwi/piRNA genome silencing defects cause immediate sterility that correlates with transposon expression and transposon-induced genomic instability. In C. elegans, mutations in the Piwi-related gene (prg-1) and other piRNA deficient mutants cause a transgenerational decline in fertility over a period of several generations. Here we show that the sterility of late generation piRNA mutants correlates poorly with increases in DNA damage signaling. Instead, sterile individuals consistently exhibit altered perinuclear germ granules. We show that disruption of germ granules does not activate transposon expression but induces multiple phenotypes found in sterile prg-1 pathway mutants. Furthermore, loss of the germ granule component pgl-1 enhances prg-1 mutant infertility. Environmental restoration of germ granule function for sterile pgl-1 mutants restores their fertility. We propose that Piwi mutant sterility is a reproductive arrest phenotype that is characterized by perturbed germ granule structure and is phenocopied by germ granule dysfunction, independent of genomic instability.
MeSH term(s)	Animals ; Animals, Genetically Modified ; Argonaute Proteins/genetics ; Atrophy ; Caenorhabditis elegans/embryology ; Caenorhabditis elegans/genetics ; Caenorhabditis elegans/physiology ; Caenorhabditis elegans Proteins/genetics ; Caenorhabditis elegans Proteins/metabolism ; DNA Damage ; Embryo, Nonmammalian ; Female ; Gene Expression Regulation ; Genomic Instability ; Germ Cells/pathology ; Infertility/genetics ; Infertility/pathology ; Larva ; Male ; Mutation ; RNA-Binding Proteins/metabolism
Chemical Substances	Argonaute Proteins ; Caenorhabditis elegans Proteins ; PGL-1 protein, C elegans ; PRG-1 protein, C elegans ; RNA-Binding Proteins
Language	English
Publishing date	2021-03-03
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2553671-0
ISSN	2041-1723 ; 2041-1723
ISSN (online)	2041-1723
ISSN	2041-1723
DOI	10.1038/s41467-021-21635-0
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Article ; Online: Germ granule dysfunction is a hallmark and mirror of Piwi mutant sterility

Maya Spichal / Bree Heestand / Katherine Kretovich Billmyre / Stephen Frenk / Craig C. Mello / Shawn Ahmed

Nature Communications, Vol 12, Iss 1, Pp 1-

2021 Volume 15

Abstract: Piwi deficiency results in sterility and is associated with transposon expression and genomic instability. Here the authors show that sterility of C. elegans Piwi prg-1 mutant is not associated with transposon-induced DNA damage but is associated with ... ...

Abstract	Piwi deficiency results in sterility and is associated with transposon expression and genomic instability. Here the authors show that sterility of C. elegans Piwi prg-1 mutant is not associated with transposon-induced DNA damage but is associated with and is phenocopied by dysfunction of germ granules.
Keywords	Science ; Q
Language	English
Publishing date	2021-03-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
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Article ; Online: Cytokinin oxidase/dehydrogenase inhibitors: outlook for selectivity and high efficiency.

Nisler, Jaroslav / Pěkná, Zuzana / Končitíková, Radka / Klimeš, Pavel / Kadlecová, Alena / Murvanidze, Nino / Werbrouck, Stefaan P O / Plačková, Lenka / Kopečný, David / Zalabák, David / Spíchal, Lukáš / Strnad, Miroslav

Journal of experimental botany

2022 Volume 73, Issue 14, Page(s) 4806–4817

Abstract: Inhibitors of cytokinin oxidase/dehydrogenase (CKX) reduce the degradation of cytokinins in plants, and this effect can be exploited in agriculture and in plant tissue culture. In this study, we examine the structure-activity relationship of two series ... ...

Abstract	Inhibitors of cytokinin oxidase/dehydrogenase (CKX) reduce the degradation of cytokinins in plants, and this effect can be exploited in agriculture and in plant tissue culture. In this study, we examine the structure-activity relationship of two series of CKX inhibitors based on diphenylurea. The compounds of Series I were derived from the recently published CKX inhibitors 3TFM-2HM and 3TFM-2HE, and we identified key substituents with increased selectivity for maize ZmCKX1 and ZmCKX4a over AtCKX2 from Arabidopsis. Series II contained compounds that further exceled in CKX inhibitory activity as well as in the ease of their synthesis. The best inhibitors exhibited half-maximal inhibitory concentration (IC50) values in low nanomolar ranges with ZmCKX1 and especially with ZmCKX4a, which is generally more resistant to inhibition. The activity of the key compounds was verified in tobacco and lobelia leaf-disk assays, where N6-isopentenyladenine was protected from degradation and promoted shoot regeneration. All the prepared compounds were further tested for toxicity against Caenorhabditis elegans, and the assays revealed clear differences in toxicity between compounds with and without a hydroxyalkyl group. In a broader perspective, this work increases our understanding of CKX inhibition and provides a more extensive portfolio of compounds suitable for agricultural and biotechnological research.
MeSH term(s)	Arabidopsis/metabolism ; Cytokinins/metabolism ; Oxidoreductases/metabolism ; Plants/metabolism ; Zea mays/metabolism
Chemical Substances	Cytokinins ; Oxidoreductases (EC 1.-) ; cytokinin oxidase (EC 1.5.99.12)
Language	English
Publishing date	2022-05-06
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2976-2
ISSN	1460-2431 ; 0022-0957
ISSN (online)	1460-2431
ISSN	0022-0957
DOI	10.1093/jxb/erac201
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Article: The adjuvant activity of two urea derivatives on cytokinins: an example of serendipitous dual effect

Brunoni, Federica / Rolli, Enrico / Polverini, Eugenia / Spíchal, Lukáš / Ricci, Ada

Plant growth regulation. 2021 Nov., v. 95, no. 2

2021

Abstract: The aim of this study was to investigate the action spectrum of two urea derivatives, the 1,3-di(benzo[d]oxazol-5-yl)urea (5-BDPU) and the 1,3-di(benzo[d]oxazol-6-yl)urea (6-BDPU). In order to evaluate a possible adjuvant activity on cytokinins the ... ...

Abstract	The aim of this study was to investigate the action spectrum of two urea derivatives, the 1,3-di(benzo[d]oxazol-5-yl)urea (5-BDPU) and the 1,3-di(benzo[d]oxazol-6-yl)urea (6-BDPU). In order to evaluate a possible adjuvant activity on cytokinins the compounds alone or in the simultaneous presence of different cytokinins were assayed either on in vitro typical cytokinin-related bioassays, or on in planta interaction with cytokinin signal transduction pathway. The compounds ability to activate the cytokinin receptor CRE1/AHK4 was studied either by a heterologous bacterial assay or by a competitive binding assay and docking simulations were performed with the crystal structure of the same receptor. Then, owing to their chemical structure which resembles that of urea-type cytokinins, the ability of 5- and 6-BDPU to inhibit the activity of cytokinin oxidase/dehydrogenase of Zea mays (ZmCKX1) was investigated and docking simulations were performed as well. Accordingly to the experimental results, we speculate that BDPUs could show a dual activity: the blocking of the conformational re-adaption of CRE1/AHK4 receptor maintaining the cytokinin inside its binding pocket, thus possibly enhancing its kinase action; the inhibition of cytokinin oxidase/dehydrogenase activity thus possibly preventing its cleavage of natural cytokinins with isoprenoid side chain.
Keywords	Zea mays ; adjuvants ; crystal structure ; cytokinins ; isoprenoids ; oxidoreductases ; plant growth ; signal transduction ; urea
Language	English
Dates of publication	2021-11
Size	p. 169-190.
Publishing place	Springer Netherlands
Document type	Article
ZDB-ID	843025-1
ISSN	1573-5087 ; 0167-6903
ISSN (online)	1573-5087
ISSN	0167-6903
DOI	10.1007/s10725-021-00735-3
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Article ; Online: The meiotic phosphatase GSP-2/PP1 promotes germline immortality and small RNA-mediated genome silencing.

Billmyre, Katherine Kretovich / Doebley, Anna-Lisa / Spichal, Maya / Heestand, Bree / Belicard, Tony / Sato-Carlton, Aya / Flibotte, Stephane / Simon, Matt / Gnazzo, Megan / Skop, Ahna / Moerman, Donald / Carlton, Peter Mark / Sarkies, Peter / Ahmed, Shawn

PLoS genetics

2019 Volume 15, Issue 3, Page(s) e1008004

Abstract: Germ cell immortality, or transgenerational maintenance of the germ line, could be promoted by mechanisms that could occur in either mitotic or meiotic germ cells. Here we report for the first time that the GSP-2 PP1/Glc7 phosphatase promotes germ cell ... ...

Abstract	Germ cell immortality, or transgenerational maintenance of the germ line, could be promoted by mechanisms that could occur in either mitotic or meiotic germ cells. Here we report for the first time that the GSP-2 PP1/Glc7 phosphatase promotes germ cell immortality. Small RNA-induced genome silencing is known to promote germ cell immortality, and we identified a separation-of-function allele of C. elegans gsp-2 that is compromised for germ cell immortality and is also defective for small RNA-induced genome silencing and meiotic but not mitotic chromosome segregation. Previous work has shown that GSP-2 is recruited to meiotic chromosomes by LAB-1, which also promoted germ cell immortality. At the generation of sterility, gsp-2 and lab-1 mutant adults displayed germline degeneration, univalents, histone methylation and histone phosphorylation defects in oocytes, phenotypes that mirror those observed in sterile small RNA-mediated genome silencing mutants. Our data suggest that a meiosis-specific function of GSP-2 ties small RNA-mediated silencing of the epigenome to germ cell immortality. We also show that transgenerational epigenomic silencing at hemizygous genetic elements requires the GSP-2 phosphatase, suggesting a functional link to small RNAs. Given that LAB-1 localizes to the interface between homologous chromosomes during pachytene, we hypothesize that small localized discontinuities at this interface could promote genomic silencing in a manner that depends on small RNAs and the GSP-2 phosphatase.
MeSH term(s)	Animals ; Caenorhabditis elegans/genetics ; Caenorhabditis elegans Proteins/genetics ; Caenorhabditis elegans Proteins/metabolism ; Chromosomal Proteins, Non-Histone/metabolism ; Chromosome Segregation ; Genome ; Germ Cells/metabolism ; Germ Cells/physiology ; Meiosis/physiology ; Meiotic Prophase I/physiology ; Methylation ; Phosphoric Monoester Hydrolases ; Protein Phosphatase 1/metabolism ; Protein Phosphatase 1/physiology ; RNA Interference/physiology ; RNA, Small Interfering
Chemical Substances	Caenorhabditis elegans Proteins ; Chromosomal Proteins, Non-Histone ; LAB-1 protein, C elegans ; RNA, Small Interfering ; Protein Phosphatase 1 (EC 3.1.3.16) ; Phosphoric Monoester Hydrolases (EC 3.1.3.2)
Language	English
Publishing date	2019-03-28
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2186725-2
ISSN	1553-7404 ; 1553-7390
ISSN (online)	1553-7404
ISSN	1553-7390
DOI	10.1371/journal.pgen.1008004
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Article ; Online: Evidence for a dual role of actin in regulating chromosome organization and dynamics in yeast.

Spichal, Maya / Brion, Alice / Herbert, Sébastien / Cournac, Axel / Marbouty, Martial / Zimmer, Christophe / Koszul, Romain / Fabre, Emmanuelle

Journal of cell science

2016 Volume 129, Issue 4, Page(s) 681–692

Abstract: Eukaryotic chromosomes undergo movements that are involved in the regulation of functional processes such as DNA repair. To better understand the origin of these movements, we used fluorescence microscopy, image analysis and chromosome conformation ... ...

Abstract	Eukaryotic chromosomes undergo movements that are involved in the regulation of functional processes such as DNA repair. To better understand the origin of these movements, we used fluorescence microscopy, image analysis and chromosome conformation capture to quantify the actin contribution to chromosome movements and interactions in budding yeast. We show that both the cytoskeletal and nuclear actin drive local chromosome movements, independently of Csm4, a putative LINC protein. Inhibition of actin polymerization reduces subtelomere dynamics, resulting in more confined territories and enrichment in subtelomeric contacts. Artificial tethering of actin to nuclear pores increased both nuclear pore complex (NPC) and subtelomere motion. Chromosome loci that were positioned away from telomeres exhibited reduced motion in the presence of an actin polymerization inhibitor but were unaffected by the lack of Csm4. We further show that actin was required for locus mobility that was induced by targeting the chromatin-remodeling protein Ino80. Correlated with this, DNA repair by homologous recombination was less efficient. Overall, interphase chromosome dynamics are modulated by the additive effects of cytoskeletal actin through forces mediated by the nuclear envelope and nuclear actin, probably through the function of actin in chromatin-remodeling complexes.
MeSH term(s)	Actins/physiology ; Chromatin/genetics ; Chromatin/metabolism ; Chromosomes, Fungal/physiology ; Membrane Proteins/physiology ; Nuclear Pore/metabolism ; Protein Multimerization ; Protein Transport ; Recombinational DNA Repair ; Saccharomyces cerevisiae/genetics ; Saccharomyces cerevisiae/metabolism ; Saccharomyces cerevisiae Proteins/physiology ; Telomere/genetics ; Telomere/metabolism
Chemical Substances	Actins ; CSM4 protein, S cerevisiae ; Chromatin ; Membrane Proteins ; Saccharomyces cerevisiae Proteins
Language	English
Publishing date	2016-02-15
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2993-2
ISSN	1477-9137 ; 0021-9533
ISSN (online)	1477-9137
ISSN	0021-9533
DOI	10.1242/jcs.175745
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Article ; Online: Cytokinin

Pokorná, Eva / Hluska, Tomáš / Galuszka, Petr / Hallmark, H Tucker / Dobrev, Petre I / Záveská Drábková, Lenka / Filipi, Tomáš / Holubová, Katarína / Plíhal, Ondřej / Rashotte, Aaron M / Filepová, Roberta / Malbeck, Jiří / Novák, Ondřej / Spíchal, Lukáš / Brzobohatý, Břetislav / Mazura, Pavel / Zahajská, Lenka / Motyka, Václav

Biomolecules

2020 Volume 11, Issue 1

Abstract: Cytokinins (CKs) are a class of phytohormones affecting many aspects of plant growth and development. In the complex process of CK homeostasis in plants, ...

Abstract	Cytokinins (CKs) are a class of phytohormones affecting many aspects of plant growth and development. In the complex process of CK homeostasis in plants,
MeSH term(s)	Arabidopsis/genetics ; Arabidopsis/metabolism ; Cytokinins/genetics ; Evolution, Molecular ; Gene Expression Regulation, Plant/genetics ; Glucosides/genetics ; Glucosyltransferases/genetics ; Oxidoreductases/genetics ; Plant Growth Regulators/genetics ; Plant Growth Regulators/metabolism ; Zea mays/genetics ; Zea mays/metabolism
Chemical Substances	Cytokinins ; Glucosides ; Plant Growth Regulators ; Oxidoreductases (EC 1.-) ; Glucosyltransferases (EC 2.4.1.-)
Language	English
Publishing date	2020-12-28
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2701262-1
ISSN	2218-273X ; 2218-273X
ISSN (online)	2218-273X
ISSN	2218-273X
DOI	10.3390/biom11010024
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Article ; Online: Design, synthesis and perception of fluorescently labeled isoprenoid cytokinins.

Kubiasová, Karolina / Mik, Václav / Nisler, Jaroslav / Hönig, Martin / Husičková, Alexandra / Spíchal, Lukáš / Pěkná, Zuzana / Šamajová, Olga / Doležal, Karel / Plíhal, Ondřej / Benková, Eva / Strnad, Miroslav / Plíhalová, Lucie

Phytochemistry

2018 Volume 150, Page(s) 1–11

Abstract: Isoprenoid cytokinins play a number of crucial roles in the regulation of plant growth and development. To study cytokinin receptor properties in plants, we designed and prepared fluorescent derivatives of 6-[(3-methylbut-2-en-1-yl)amino]purine ( ... ...

Abstract	Isoprenoid cytokinins play a number of crucial roles in the regulation of plant growth and development. To study cytokinin receptor properties in plants, we designed and prepared fluorescent derivatives of 6-[(3-methylbut-2-en-1-yl)amino]purine (N
MeSH term(s)	4-Chloro-7-nitrobenzofurazan/pharmacology ; Adenine/analogs & derivatives ; Adenine/chemistry ; Arabidopsis/metabolism ; Arabidopsis Proteins/metabolism ; Carbocyanines/chemistry ; Coloring Agents/chemistry ; Cytokinins/chemistry ; Cytokinins/pharmacology ; Fluorescent Dyes/chemistry ; Gene Expression Regulation, Plant ; Isopentenyladenosine/chemical synthesis ; Isopentenyladenosine/chemistry ; Isopentenyladenosine/pharmacology ; Microscopy, Confocal ; Molecular Structure ; Plant Development ; Plant Growth Regulators/metabolism ; Purines/chemistry ; Receptors, Cytokine/antagonists & inhibitors ; Receptors, Cytokine/chemistry ; Rhodamines/chemistry ; Seedlings/metabolism ; Terpenes/metabolism ; Zea mays/metabolism
Chemical Substances	6-((3-methylbut-2-en-1-yl)amino)purine ; Arabidopsis Proteins ; Carbocyanines ; Coloring Agents ; Cytokinins ; Fluorescent Dyes ; Plant Growth Regulators ; Purines ; Receptors, Cytokine ; Rhodamines ; Terpenes ; cyanine dye 5 ; N(6)-(delta(2)-isopentenyl)adenine (2365-40-4) ; N(6),N(6)-dimethyladenine (649SA4S2CV) ; Isopentenyladenosine (7724-76-7) ; 4-Chloro-7-nitrobenzofurazan (EQF2794IRE) ; Adenine (JAC85A2161) ; rhodamine B (K7G5SCF8IL) ; purine (W60KTZ3IZY)
Language	English
Publishing date	2018-06
Publishing country	England
Document type	Journal Article
ZDB-ID	208884-8
ISSN	1873-3700 ; 0031-9422
ISSN (online)	1873-3700
ISSN	0031-9422
DOI	10.1016/j.phytochem.2018.02.015
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Article: C2-substituted aromatic cytokinin sugar conjugates delay the onset of senescence by maintaining the activity of the photosynthetic apparatus

Vylíčilová, Hana / Alexandra Husičková / Josef Srovnal / Karel Doležal / Lucie Plíhalová / Lukáš Spíchal / Ondřej Plíhal

Phytochemistry. 2016 Feb., v. 122

2016

Abstract: Cytokinins are plant hormones with biological functions ranging from coordination of plant growth and development to the regulation of senescence. A series of 2-chloro-N6-(halogenobenzylamino)purine ribosides was prepared and tested for cytokinin ... ...

Abstract	Cytokinins are plant hormones with biological functions ranging from coordination of plant growth and development to the regulation of senescence. A series of 2-chloro-N6-(halogenobenzylamino)purine ribosides was prepared and tested for cytokinin activity in detached wheat leaf senescence, tobacco callus and Amaranthus bioassays. The synthetic compounds showed significant activity, especially in delaying senescence in detached wheat leaves. They were also tested in bacterial receptor bioassays using both monocot and dicot members of the cytokinin receptor family. Most of the derivatives did not trigger cytokinin signaling via the AHK3 and AHK4 receptors from Arabidopsis thaliana in the bacterial assay, but some of them specifically activated the ZmHK1 receptor from Zea mays and were also more active than the aromatic cytokinin BAP in an ARR5::GUS cytokinin bioassay using transgenic Arabidopsis plants. Whole transcript expression analysis was performed using an Arabidopsis model to gather information about the reprogramming of gene transcription when senescent leaves were treated with selected C2-substituted aromatic cytokinin ribosides. Genome-wide expression profiling revealed that the synthetic halogenated derivatives induced the expression of genes related to cytokinin signaling and metabolism. They also prompted both up- and down-regulation of a unique combination of genes coding for components of the photosystem II (PSII) reaction center, light-harvesting complex II (LHCII), and the oxygen-evolving complex, as well as several stress factors responsible for regulating photosynthesis and chlorophyll degradation. Chlorophyll content and fluorescence analyses demonstrated that treatment with the halogenated derivatives increased the efficiency of PSII photochemistry and the abundance of LHCII relative to DMSO- and BAP-treated controls. These findings demonstrate that it is possible to manipulate and fine-tune leaf longevity using synthetic aromatic cytokinin analogs.
Keywords	Amaranthus ; Arabidopsis thaliana ; bioassays ; callus ; chlorophyll ; cytokinins ; dimethyl sulfoxide ; fluorescence ; gene expression ; gene expression regulation ; genes ; growth and development ; leaves ; light harvesting complex ; longevity ; models ; oxygen evolving complex ; photochemistry ; plant growth ; plant hormones ; receptors ; senescence (aging) ; sugars ; tobacco ; transcription (genetics) ; transcriptomics ; transgenic plants ; wheat ; Zea mays
Language	English
Dates of publication	2016-02
Size	p. 22-33.
Publishing place	Elsevier Ltd
Document type	Article
ZDB-ID	208884-8
ISSN	1873-3700 ; 0031-9422
ISSN (online)	1873-3700
ISSN	0031-9422
DOI	10.1016/j.phytochem.2015.12.001
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