Biology subjects

Hause, G.

Publications and source records attributed to Hause, G..

6 recordsLinked to original sources

bioRxiv2026

The role of MYB21 in Arabidopsis: transcending flower-specific functions to vegetative tissue

Jasmonic acid (JA) and its derivatives are lipid-derived phytohormones that coordinate plant growth, development, and stress responses through the bioactive conjugate jasmonoyl-isoleucine (JA-Ile). Their role in reproductive development is well established, particularly in stamen maturation through the R2R3-MYB transcription factor MYB21, which has been considered largely flower-specific. Here, we reveal a previously unrecognized role of MYB21 in vegetative tissues of Arabidopsis thaliana. Although basal MYB21 transcript levels in leaves are extremely low and spatially restricted, wounding and exogenous hormone applications induced MYB21 transcription in a JA- and COI1-dependent manner. Transcriptional GUS reporter analyses showed localized MYB21 promoter activity in specialized epidermal cells, including trichomes, hydathodes, and in the vasculature at wound sites. Functional characterization using the myb21-5 mutant indicated roles in germination and vegetative growth, partially phenocopying JA-insensitive mutants despite unaltered JA biosynthesis and signaling. Transcriptome profiling further revealed changes in expression of genes involved in lignin biosynthesis, light-harvesting complex components, cytokinin pathways, and defense-related responses, consistent with reduced resistance of myb21-5 to insect herbivory and infection by Botrytis cinerea. Together, these findings identify MYB21 as a JA-responsive regulator of growth and defense in seedlings and leaves, extending its function beyond reproductive development. HighlightThe transcription factor MYB21, previously considered flower-specific, is induced by jasmonate signaling in Arabidopsis leaves and seedlings, where it regulates growth and defense responses, thereby expanding its role beyond reproductive development.

plant biology↗

bioRxiv2023

Stromule geometry allows optimal spatial regulation of organelle interactions in the quasi-2D cytoplasm

In plant cells, plastids form elongated extensions called stromules, the regulation and purposes of which remain unclear. Here we quantitatively explore how different stromule structures serve to enhance the ability of a plastid to interact with other organelles: increasing the effective space for interaction and biomolecular exchange between organelles. Strikingly, electron microscopy and confocal imaging showed that the cytoplasm in Arabidopsis thaliana and Nicotiana benthamiana epidermal cells is extremely thin (around 100 nm in regions without organelles), meaning that inter-organelle interactions effectively take place in 2D. We combine these imaging modalities with mathematical modelling and new in planta experiments to demonstrate how different the elongation of stromules (single or multiple, linear or branching) could be employed to optimise different aspects of inter-organelle interaction capacity in this 2D space. Stromule formation and branching is shown to provide a proportionally higher benefit to interaction capacity in 2D than in 3D. Additionally we find this benefit depends on optimal plastid spacing. We hypothesize that cells can promote the formation of different stromule architectures in the quasi-2D cytoplasm to optimise their interaction interface to meet specific requirements. These results provide new insight into the mechanisms underlying the transition from low to high stromule numbers during biotic stress, the consequences for interaction with smaller organelles, how plastid access and plastid to nucleus signalling is balanced, as well as the impact of plastid density on organelle interaction.

plant biology↗

bioRxiv2023

Preparation of physiologically active inside-out vesicles from plant inner mitochondrial membranes

For many metabolites, the major barrier between cytosol and mitochondrial matrix is the inner envelope membrane of mitochondria, the site of the respiratory electron transport chain. In consequence, it houses numerous transporters which facilitate the controlled exchange of metabolites, ions, and even proteins between these cellular compartments. While their import into the organelle can be studied with isolated mitochondria or mitoplasts, the analysis of their export from the matrix into the intermembrane space or even the cytosol demands for more sophisticated approaches. Among those, inside-out inner membrane vesicles are particularly useful, since they allow the direct presentation of the potential export substrates to the membrane without prior import into the organelle. Here we present a protocol for the isolation of such inside-out vesicles of the inner envelope membrane of plant mitochondria based on repeated freeze/thaw-cycles of freshly prepared mitoplasts. Electron microscopy and Western analysis could show that the majority of the vesicles have single envelope membranes in an inside-out topology. The vesicles are furthermore physiologically active, as demonstrated by assays measuring the enzymatic activities of Complex I (NADH dehydrogenase), Complex V (ATP synthase) and the mitochondrial processing peptidase (MPP) associated with Complex III. Hence, the method presented here provides a good basis for further studies of the inner mitochondrial envelope membrane and mitochondrial export processes.

cell biology↗

bioRxiv2022

AI-guided cryo-EM probes a thermophilic cell-free system with succinyl-coA manufacturing capability

Cell-free systems display tremendous potential for biotechnological applications, complementing in vitro reconstituted enzymatic processes and traditional expression systems. However, they often represent "black boxes" without much insight into their components. Here, we characterize a thermophilic cell-free system that produces succinyl-CoA and discern its intrinsic, non-stochastic organization. By employing biochemical, biophysical, and bioinformatic methods we resolve its molecular composition, 3D architecture and molecular function at atomic resolution. We further report the high-resolution cryo-EM structure of the reactions main component, the oxoglutarate dehydrogenase complex core (E2o), which displays various structural adaptations. These include hydrogen bonding patterns confining interactions of participating enzymes (E1o-E2o-E3), electrostatic tunneling that drives inter-communication between subunits, and the presence of a flexible subunit, the E3BPo connecting E2o and E3. This multi-scale analysis of a cell-free system provides a blueprint for structure-function studies of complex mixtures of biotechnological value.

biochemistry↗

bioRxiv2022

Delineating organizational principles of the endogenous L-A virus by cryo-EM and computational analysis of native cell extracts

The high abundance of most viruses in infected host cells benefits their structural characterization; endogenous viruses are present in low copy numbers, however, and are therefore challenging to investigate. Here, we retrieve cell extracts enriched with an endogenous virus, the yeast L-A virus. The determined cryo-EM structure discloses capsid-stabilizing cation-{pi} stacking and an interplay of non-covalent interactions from ten distinct capsomere interfaces. The capsid-embedded mRNA decapping active site trench is supported by a constricting movement of two opposite-facing loops. tRNA-loaded polysomes and other biomacromolecules, presumably mRNA, are found in virus proximity while stacked dsRNA bundles and the sub-stoichiometric polymerase localize underneath the capsid surface. Mature viruses participate in larger viral communities resembling their rare in-cell equivalents in terms of size, composition, and inter-virus distances. Our results collectively describe a 3D-architecture of a viral milieu, opening the door to cellextract-based high-resolution structural virology.

biochemistry↗

bioRxiv2018

Phosphorylation of the exocyst subunit Exo70B2 contributes to the regulation of its function

The exocyst is a conserved hetero-octameric complex that mediates early tethering of post-Golgi vesicles during exocytosis. Its Exo70 subunit functions as a spatiotemporal regulator by mediating numerous interactions with proteins and lipids. However, a molecular understanding of the exocyst functions remains challenging. Exo70B2 localized to dynamic foci at the plasma membrane and transited through Brefeldin A (BFA)-sensitive compartments, indicating that it participates in conventional secretion. Conversely, treatment with the immunogenic peptide flg22 or the salicylic acid (SA) defence hormone analogue Benzothiadiazole (BTH), induced Exo70B2 transport into the vacuole where it colocalized with autophagic markers AUTOPHAGY-RELATED PROTEIN 8 (ATG8) and NEIGHBOR OF BRCA1 GENE 1 (NBR1). According with its role in immunity, we discovered that Exo70B2 interacts with and is phosphorylated by the MITOGEN-ACTIVATED PROTEIN KINASE 3 (MPK3). Mimicking phosphorylation inhibited Exo70B2 localization at sites of active secretion. By contrast, lines expressing phosphonull variants displayed higher Effector-Triggered Immunity and were hypersensitive to BTH, conditions known to induce the secretory pathway. Our results suggest a molecular mechanism by which phosphorylation of Exo70B2 regulates interaction with the plasma membrane, and couples the secretory pathway with cellular signalling.

cell biology↗