TY - THES A1 - Schumacher, Julia T1 - Regulation and function of STERILE APETALA in Arabidopsis flower development N2 - STERILE APETALA (SAP) is known to be an essential regulator of flower development for over 20 years. Loss of SAP function in the model plant Arabidopsis thaliana is associated with a reduction of floral organ number, size and fertility. In accordance with the function of SAP during early flower development, its spatial expression in flowers is confined to meristematic stages and to developing ovules. However, to date, despite extensive research, the molecular function of SAP and the regulation of its spatio-temporal expression still remain elusive. In this work, amino acid sequence analysis and homology modeling revealed that SAP belongs to the rare class of plant F-box proteins with C-terminal WD40 repeats. In opisthokonts, this type of F-box proteins constitutes the substrate binding subunit of SCF complexes, which catalyze the ubiquitination of proteins to initiate their proteasomal degradation. With LC-MS/MS-based protein complex isolation, the interaction of SAP with major SCF complex subunits was confirmed. Additionally, candidate substrate proteins, such as the growth repressor PEAPOD 1 and 2 (PPD1/2), could be revealed during early stages of flower development. Also INDOLE-3-BUTYRIC ACID RESPONSE 5 (IBR5) was identified among putative interactors. Genetic analyses indicated that, different from substrate proteins, IBR5 is required for SAP function. Protein complex isolation together with transcriptome profiling emphasized that the SCFSAP complex integrates multiple biological processes, such as proliferative growth, vascular development, hormonal signaling and reproduction. Phenotypic analysis of sap mutant and SAP overexpressing plants positively correlated SAP function with plant growth during reproductive and vegetative development. Furthermore, to elaborate on the transcriptional regulation of SAP, publicly available ChIP-seq data of key floral homeotic proteins were reanalyzed. Here, it was shown that the MADS-domain transcription factors APETALA 1 (AP1), APETALA 3 (AP3), PISTILLATA (PI), AGAMOUS (AG) and SEPALLATA 3 (SEP3) bind to the SAP locus, which indicates that SAP is expressed in a floral organ-specific manner. Reporter gene analyses in combination with CRISPR/Cas9-mediated deletion of putative regulatory regions further demonstrated that the intron contains major regulatory elements of SAP in Arabidopsis thaliana. In conclusion, these data indicate that SAP is a pleiotropic developmental regulator that acts through tissue-specific destabilization of proteins. The presumed transcriptional regulation of SAP by the floral MADS-domain transcription factors could provide a missing link between the specification of floral organ identity and floral organ growth pathways. KW - STERILE APETALA KW - SAP KW - flower development KW - organ size KW - F-box KW - WD40 KW - SCF complex KW - ubiquitin KW - proteasomal degradation KW - MADS Y1 - 2019 ER - TY - JOUR A1 - Johnson, Kim L. A1 - Lenhard, Michael T1 - Genetic control of plant organ growth JF - New phytologist : international journal of plant science N2 - The growth of plant organs is under genetic control. Work in model species has identified a considerable number of genes that regulate different aspects of organ growth. This has led to an increasingly detailed knowledge about how the basic cellular processes underlying organ growth are controlled, and which factors determine when proliferation gives way to expansion, with this transition emerging as a critical decision point during primordium growth. Progress has been made in elucidating the genetic basis of allometric growth and the role of tissue polarity in shaping organs. We are also beginning to understand how the mechanisms that determine organ identity influence local growth behaviour to generate organs with characteristic sizes and shapes. Lastly, growth needs to be coordinated at several levels, for example between different cell layers and different regions within one organ, and the genetic basis for such coordination is being elucidated. However, despite these impressive advances, a number of basic questions are still not fully answered, for example, whether and how a growing primordium keeps track of its size. Answering these questions will likely depend on including additional approaches that are gaining in power and popularity, such as combined live imaging and modelling. KW - growth coordination KW - organ growth KW - organ identity KW - organ shape KW - organ size Y1 - 2011 U6 - https://doi.org/10.1111/j.1469-8137.2011.03737.x SN - 0028-646X VL - 191 IS - 2 SP - 319 EP - 333 PB - Wiley-Blackwell CY - Malden ER - TY - JOUR A1 - Nguyen, Hung M. A1 - Schippers, Jos H. M. A1 - Goni-Ramos, Oscar A1 - Christoph, Mathias P. A1 - Dortay, Hakan A1 - van der Hoorn, Renier A. L. A1 - Müller-Röber, Bernd T1 - An upstream regulator of the 26S proteasome modulates organ size in Arabidopsis thaliana JF - The plant journal N2 - In both animal and plant kingdoms, body size is a fundamental but still poorly understood attribute of biological systems. Here we report that the Arabidopsis NAC transcription factor Regulator of Proteasomal Gene Expression' (RPX) controls leaf size by positively modulating proteasome activity. We further show that the cis-element recognized by RPX is evolutionarily conserved between higher plant species. Upon over-expression of RPX, plants exhibit reduced growth, which may be reversed by a low concentration of the pharmacological proteasome inhibitor MG132. These data suggest that the rate of protein turnover during growth is a critical parameter for determining final organ size. KW - Arabidopsis thaliana KW - organ size KW - evolution KW - leaf development KW - proteasome KW - gene regulatory network Y1 - 2013 U6 - https://doi.org/10.1111/tpj.12097 SN - 0960-7412 VL - 74 IS - 1 SP - 25 EP - 36 PB - Wiley-Blackwell CY - Hoboken ER - TY - THES A1 - Trost, Gerda T1 - Poly(A) Polymerase 1 (PAPS1) influences organ size and pathogen response in Arabidopsis thaliana T1 - Poly(A) Polymerase 1 (PAPS1) beeinflusst die Organgröße und Pathogenantwort in Arabidopsis thaliana N2 - Polyadenylation of pre-mRNAs is critical for efficient nuclear export, stability, and translation of the mature mRNAs, and thus for gene expression. The bulk of pre-mRNAs are processed by canonical nuclear poly(A) polymerase (PAPS). Both vertebrate and higher-plant genomes encode more than one isoform of this enzyme, and these are coexpressed in different tissues. However, in neither case is it known whether the isoforms fulfill different functions or polyadenylate distinct subsets of pre-mRNAs. This thesis shows that the three canonical nuclear PAPS isoforms in Arabidopsis are functionally specialized owing to their evolutionarily divergent C-terminal domains. A moderate loss-of-function mutant in PAPS1 leads to increase in floral organ size, whereas leaf size is reduced. A strong loss-of-function mutation causes a male gametophytic defect, whereas a weak allele leads to reduced leaf growth. By contrast, plants lacking both PAPS2 and PAPS4 function are viable with wild-type leaf growth. Polyadenylation of SMALL AUXIN UP RNA (SAUR) mRNAs depends specifically on PAPS1 function. The resulting reduction in SAUR activity in paps1 mutants contributes to their reduced leaf growth, providing a causal link between polyadenylation of specific pre-mRNAs by a particular PAPS isoform and plant growth. Additionally, opposite effects of PAPS1 on leaf and flower growth reflect the different identities of these organs. The overgrowth of paps1 mutant petals is due to increased recruitment of founder cells into early organ primordia whereas the reduced leaf size is due to an ectopic pathogen response. This constitutive immune response leads to increased resistance to the biotrophic oomycete Hyaloperonospora arabidopsidis and reflects activation of the salicylic acid-independent signalling pathway downstream of ENHANCED DISEASE SUSCEPTIBILITY1 (EDS1)/PHYTOALEXIN DEFICIENT4 (PAD4). Immune responses are accompanied by intracellular redox changes. Consistent with this, the redox-status of the chloroplast is altered in paps1-1 mutants. The molecular effects of the paps1-1 mutation were analysed using an RNA sequencing approach that distinguishes between long- and short tailed mRNA. The results shown here suggest the existence of an additional layer of regulation in plants and possibly vertebrate gene expression, whereby the relative activities of canonical nuclear PAPS isoforms control de novo synthesized poly(A) tail length and hence expression of specific subsets of mRNAs. N2 - Polyadenylierung von prä-mRNAs ist entscheidend für den Export aus dem Zellkern, die Stabilität und die Translation der reifen mRNAs und dadurch für die Genexpression. Der Großteil der mRNAs wird durch sogenannte canonische Poly(A) Polymerasen (cPAPS) prozessiert. Die Genome von sowohl Wirbeltieren als auch Pflanzen kodieren mehr als eine Isoform dieser Enzyme, welche gleichzeitig in verschiedenen Geweben exprimiert werden. Es ist jedoch kein Beispiel bekannt, das zeigt, ob die verschiedenen Isoformen unterschiedliche Funktionen einnehmen bzw. verschiedene Untergruppen von mRNAs polyadenylieren. Diese Arbeit zeigt, dass drei canonische PAPS Isoformen in Arabidopsis thaliana aufgrund ihrer evolutionär unterschiedlichen C-terminalen Domänen spezialisierte Funktionen haben. Eine schwache Verlust-Mutation im PAPS1 Gen bewirkt eine Vergrößerung der Blütenorgane, während die Blattgröße vermindert ist. Eine starke Verlust-Mutation bewirkt zusätzlich einen Defekt der männlichen Keimzellen. Im Gegenzug dazu sind Mutanten des PAPS2 oder PAPS4 Gens gesund und zeigen ein normales Wachstum. Polyadenylierung von SMALL AUXIN UP RNA (SAUR) mRNAs hängt spezifisch von der Funktion von PAPS1 ab. Die daraus entstehende Reduzierung der SAUR Aktivität in den paps1 Mutanten trägt zur Verringerung der Blattgröße bei und stellt eine kausale Verbindung zwischen Polyadenylierung spezifischer mRNAs durch bestimmte PAPS Isoformen und Pflanzenwachstum dar. Zusätzlich spiegeln die unterschiedlichen Effekte von PAPS1 auf Blüten und Blätter die Identitäten dieser Organe wieder. Das übermäßige Wachstum der mutanten Petalen beruht auf einer erhöhten Anzahl an Gründer-Zellen im frühen Primordium, wohingegen die verminderte Blattgröße auf eine ektopische Pathogen Antwort zurückzuführen ist. Diese konstitutive Immunantwort bewirkt eine erhöhte Resistenz der Mutanten gegenüber dem biotrophen Oomyceten Hyaloperonospora arabidopsidis und reflektiert die Aktivierung des Salizylsäure unabhängigen Signalweges von ENHANCED DISEASE SUSCEPTIBILITY1 (EDS1)/PHYTOALEXIN DEFICIENT4 (PAD4). Immunantworten sind von Veränderungen des intrazellulären Redoxpotenzials gekennzeichnet. Damit übereinstimmend zeigen die Chloroplasten der paps1-1 Mutanten ein verändertes Redoxpotenzial. Zur genaueren Aufklärung der molekularen Effekte der paps1 1 mutation wurde eine RNA-Sequenzierungsmethode verwendet, die zwischen mRNAs mit langem oder kurzem Poly(A) Schwanz unterscheidet. Die Aktivitäten der verschiedenen canonischen PAPS Isoformen kontrollieren die Länge des neu synthetisierten poly(A) Schwanzes und damit die Expression spezifischer Untergruppen von mRNAs. Dadurch lassen die hier gezeigten Ergebnisse eine weitere Ebene der Genregulierung in Pflanzen, und möglicherweise auch in anderen Eukaryoten, vermuten. KW - Polyadenylierung KW - Pathogenantwort KW - Organgröße KW - polyadenylation KW - pathogen response KW - organ size Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-72345 ER -