Plant stress : hitting pause on the cell cycle

Two recently discovered transcription factors stop cells from dividing when plants face extreme heat and DNA damage

New insights into the mechanisms of phytochrome-cryptochrome coaction.

Contents Summary 547 I. Introduction 547 II. Phytochromes mediate light-induced transcription of BICs to inactivate cryptochromes 548 III. PPKs phosphorylate light-signaling proteins and histones to affect plant development 548 IV. Prospect 550 Acknowledgements 550 References 550 SUMMARY: Plants perceive and respond to light signals by multiple sensory photoreceptors, including phytochromes and cryptochromes, which absorb different wavelengths of light to regulate genome expression and plant development. Photophysiological analyses have long revealed the coordinated actions of different photoreceptors, a phenomenon referred to as the photoreceptor coaction. The mechanistic explanations of photoreceptor coactions are not fully understood. The function of direct protein-protein interaction of phytochromes and cryptochromes and common signaling molecules of these photoreceptors, such as SPA1/COP1 E3 ubiquitin ligase complex and bHLH transcription factors PIFs, would partially explain phytochrome-cryptochrome coactions. In addition, newly discovered proteins that block cryptochrome photodimerization or catalyze cryptochrome phosphorylation may also participate in the phytochrome and cryptochrome coaction. This Tansley insight, which is not intended to make a comprehensive review of the studies of photoreceptor coactions, attempts to highlight those recent findings and their possible roles in the photoreceptor coaction

The UBP5 histone H2A deubiquitinase counteracts PRCs-mediated repression to regulate Arabidopsis development

Polycomb Repressive Complexes (PRCs) control gene expression through the incorporation of H2Aub and H3K27me3. In recent years, there is increasing evidence of the complexity of PRCs’ interaction networks and the interplay of these interactors with PRCs in epigenome reshaping, which is fundamental to understand gene regulatory mechanisms. Here, we identified UBIQUITIN SPECIFIC PROTEASE 5 (UBP5) as a chromatin player able to counteract the deposition of the two PRCs’ epigenetic hallmarks in Arabidopsis thaliana. We demonstrated that UBP5 is a plant developmental regulator based on functional analyses of ubp5-CRISPR Cas9 mutant plants. UBP5 promotes H2A monoubiquitination erasure, leading to transcriptional de-repression. Furthermore, preferential association of UBP5 at PRC2 recruiting motifs and local H3K27me3 gaining in ubp5 mutant plants suggest the existence of functional interplays between UBP5 and PRC2 in regulating epigenome dynamics. In summary, acting as an antagonist of the pivotal epigenetic repressive marks H2Aub and H3K27me3, UBP5 provides novel insights to disentangle the complex regulation of PRCs’ activities

Arabidopsis S2Lb links AtCOMPASS-like and SDG2 activity in H3K4me3 independently from histone H2B monoubiquitination.

The functional determinants of H3K4me3, their potential dependency on histone H2B monoubiquitination, and their contribution to defining transcriptional regimes are poorly defined in plant systems. Unlike in Saccharomyces cerevisiae, where a single SET1 protein catalyzes H3K4me3 as part of COMPlex of proteins ASsociated with Set1 (COMPASS), in Arabidopsis thaliana, this activity involves multiple histone methyltransferases. Among these, the plant-specific SET DOMAIN GROUP 2 (SDG2) has a prominent role. We report that SDG2 co-regulates hundreds of genes with SWD2-like b (S2Lb), a plant ortholog of the Swd2 axillary subunit of yeast COMPASS. We show that S2Lb co-purifies with the AtCOMPASS core subunit WDR5, and both S2Lb and SDG2 directly influence H3K4me3 enrichment over highly transcribed genes. S2Lb knockout triggers pleiotropic developmental phenotypes at the vegetative and reproductive stages, including reduced fertility and seed dormancy. However, s2lb seedlings display little transcriptomic defects as compared to the large repertoire of genes targeted by S2Lb, SDG2, or H3K4me3, suggesting that H3K4me3 enrichment is important for optimal gene induction during cellular transitions rather than for determining on/off transcriptional status. Moreover, unlike in budding yeast, most of the S2Lb and H3K4me3 genomic distribution does not rely on a trans-histone crosstalk with histone H2B monoubiquitination. Collectively, this study unveils that the evolutionarily conserved COMPASS-like complex has been co-opted by the plant-specific SDG2 histone methyltransferase and mediates H3K4me3 deposition through an H2B monoubiquitination-independent pathway in Arabidopsis

DET1-mediated degradation of a SAGA-like deubiquitination module controls H2Bub homeostasis

DE-ETIOLATED 1 (DET1) is an evolutionarily conserved component of the ubiquitination machinery that mediates the destabilization of key regulators of cell differentiation and proliferation in multicellular organisms. In this study, we provide evidence from Arabidopsis that DET1 is essential for the regulation of histone H2B monoubiquitination (H2Bub) over most genes by controlling the stability of a deubiquitination module (DUBm). In contrast with yeast and metazoan DUB modules that are associated with the large SAGA complex, the Arabidopsis DUBm only comprises three proteins (hereafter named SGF11, ENY2 and UBP22) and appears to act independently as a major H2Bub deubiquitinase activity. Our study further unveils that DET1-DDB1-Associated-1 (DDA1) protein interacts with SGF11 in vivo, linking the DET1 complex to light-dependent ubiquitin-mediated proteolytic degradation of the DUBm. Collectively, these findings uncover a signaling path controlling DUBm availability, potentially adjusting H2Bub turnover capacity to the cell transcriptional status

Chromatin dynamics during interphase and cell division:similarities and differences between model and crop plants

Genetic information in the cell nucleus controls organismal development, responses to the environment and finally ensures own transmission to the next generations. To achieve so many different tasks, the genetic information is associated with structural and regulatory proteins, which orchestrate nuclear functions in time and space. Furthermore, plant life strategies require chromatin plasticity to allow a rapid adaptation to abiotic and biotic stresses. Here, we summarize current knowledge on the organisation of plant chromatin and dynamics of chromosomes during interphase and mitotic and meiotic cell divisions for model and crop plants differing as to the genome size, ploidy and amount of genomic resources available. The existing data indicate that chromatin changes accompany most (if not all) cellular processes and that there are both shared and unique themes in the chromatin structure and global chromosome dynamics among species. Ongoing efforts to understand the molecular mechanisms involved in chromatin organisation and remodeling have, together with the latest genome editing tools, potential to unlock crop genomes for innovative breeding strategies and improvements of various traits

Chromatin dynamics during photomorphogenesis in Arabidopsis thaliana

Les états chromatiniens peuvent être étudiés à l’échelle des unités transcriptionnelles par des approches moléculaires ou à l'échelle plus globale de l'hétérochromatine structurée au sein de chromocentres par des approches cytogénétiques. Ces deux niveaux d’organisation de la chromatine sont dynamiques et influencent l'ensemble des processus nucléaires. L’objectif de cette thèse était d'avancer la compréhension des dynamiques chromatiniennes à ces deux échelles chez la plante modèle Arabidopsis thaliana, en se focalisant sur une transition développementale majeure, la photomorphogenèse. Le processus de dé-étiolement implique la reprogrammation de l’expression de centaines de gènes en réponse à la lumière, constituant ainsi un excellent modèle d'étude. La première partie des travaux montre que la reprogrammation de l’expression du génome au cours de la photomorphogenèse est associée à des dynamiques de l’hétérochromatine qui sont régulés de façon différentielles dans les hypocotyles et les cotylédons. Ces dynamiques à grande échelle ont des conséquences localement, car les états décompactés sont associés à la réactivation d'éléments hétérochromatiniens répétés. Dans une deuxième partie, le répresseur transcriptionnel DE-ETIOLATED-1 (DET1) a été utilisé afin de rechercher l'implication de régulateurs de la photomorphogenèse dans les mécanismes chromatiniens. Ce répresseur majeur de la photomorphogenèse peut lier l'histone H2B et influence le niveau global de sa modification par mono-ubiquitination (H2Bub). Dans le cadre de ma thèse, j'ai révélé d'une part l’existence d’interactions génétiques entre DET1 et les gènes contrôlant l’homéostasie de H2Bub et d'autre part un défaut de la régulation chromatinienne des variants des gènes ribosomiques 5S et 45S dans le mutant det1-1. L’ensemble de ces données permet de proposer un modèle impliquant DET1 dans la régulation de H2Bub de façon différentielle dans l’euchromatine et l’hétérochromatine, constituant ainsi le premier lien entre régulateurs de la photomorphogenèse et modifications des histones. La marque H2Bub étant directement liée à l'activité transcriptionnelle chez divers eucaryotes, l'impact de H2Bub sur l'expression des gènes durant la photomorphogenèse a été analysé. La combinaison d’approches épigénomiques et transcriptomiques a permis de montrer que le gain de H2Bub est associé à l’induction des gènes. L’utilisation du mutant hub1 dans lequel le dépôt de H2Bub est aboli a également permis de révéler le rôle de cette marque pour une régulation rapide de l’induction et de la répression de nombreux gènes. De façon générale, ce travail a révélé des dynamiques chromatiniennes impliquant des réorganisations massives au niveau cytologique ainsi que des variations fines des modifications d'histones au niveau des gènes de l'euchromatine, ainsi que le rôle de DET1 dans la régulation de ces processus. Il ouvre donc la voie à l'étude des connections entre ces deux échelles de dynamiques pour la régulation de l'activité transcriptionnelle, liant compartimentation nucléaire et activité des gènes dans le contexte global de la réponse aux signaux lumineux.Chromatin states can be studied both at the level of individual transcriptional units by molecular approaches or at the larger scale of heterochromatin by cytogenetic approaches. These two levels of chromatin organization are dynamic and influence all nuclear processes. The objective was to enhance the understanding of chromatin dynamics at these two scales in the model plant Arabidopsis thaliana, focusing on a major developmental transition, photomorphogenesis. The process of de-etiolation involves the reprogramming of the expression of hundreds of genes in response to the perception of light therefore constituting an excellent experimental system. The first part of the work shows that reprogramming of genome expression during photomorphogenesis is associated with heterochromatin dynamics that is differentially regulated in the hypocotyls and the cotyledons. These widespread dynamics have local consequences, as the decompacted states are associated with reactivation of heterochromatic repeat elements. In the second part, the transcriptional repressor DE-ETIOLATED-1 (DET1) was used to investigate the involvement of photomorphogenesis regulators in chromatin mechanisms. This major repressor of photomorphogenesis can bind histone H2B and influences the overall level of mono-ubiquitinated H2B (H2Bub). As part of my thesis, I uncovered the existence of genetic interactions between DET1 and the genes controlling H2Bub homeostasis and also a defect in the regulation of the chromatin around the 45S and 5S ribosomal genes in the mutant det1-1. These data have led me to propose a model involving DET1 in the differential regulation of H2Bub in heterochromatin and euchromatin, thus constituting for the first time a link between photomorphogenesis regulators and histone modifications. Because the H2Bub mark has been directly linked to transcriptional activity in a diverse range of eukaryotes, I analysed the impact of H2Bub on gene expression during photomorphogenesis in the third part of my thesis. The combination of transcriptomic and epigenomic approaches showed that the gain of H2Bub is associated with gene induction. The use of a hub1 mutant in which H2Bub deposition is abolished also revealed the role of this mark for the rapid control of many genes. In general terms, this work has revealed both dynamic chromatin changes that result in major genome reorganizations at the cytological scale and fine variations of histone modifications on euchromatic genes, as well as the role of DET1 in regulating these changes. My study paves the way for further studies on the connections between these two scales of dynamics and their function in the nuclear localization and changes in expression of genes in the overall context of light signaling