619 research outputs found

    Dynamique de la paroi cellulaire dans la régulation de la morphogenèse et de la croissance cellulaire

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    Cells in nature develop in a wide range of forms, following diverse growth patterns. Despite the importance of these fundamental processes, how cells regulate their growth and morphogenesis is still poorly understood. In this thesis, I explored these processes, focusing my investigations on tip growing walled cells and in particular, by exploiting the fission yeast Schyzosaccharomyces pombe, adopting a mainly biomechanical approach. To this aim, I first developed novel methods to measure key cell wall mechanical parameters in vivo and in large scale, which allowed the very first observations of cell wall dynamics. This revealed that the cell wall is softer and highly variable at growing poles, and almost stable and stiffer at non-growing sites. During elongation, there is an interplay between wall mechanics and cell growth, whose active control allows cell expansion while preserving cell integrity. In addition, I observed that there is a strong correlation between cell wall mechanics and cell morphology, and ectopic perturbations of wall properties directly affect shape establishment and maintenance. Together my results show that the regulation of wall mechanics is fundamental in the determination of cell dynamics in tip growing walled cells. Moreover, this suggests that dynamic observation of cell surface mechanics is crucial for a complete understanding of multifactorial and complex processes as growth and morphogenesis.Les cellules dans la nature se développent dans un large éventail de formes, suivant divers modèles de croissance. Malgré l'importance de ces processus fondamentaux, la façon dont les cellules régulent leur croissance et leur morphogenèse est encore mal comprise. Dans cette thèse, j'ai exploré ces aspects, avec une approche principalement biomécanique, en concentrant mes investigations sur des cellules à paroi à croissance de pointe et en exploitant en particulier la levure fissipare Schyzosaccharomyces pombe. J'ai d'abord développé de nouvelles méthodes pour mesurer les paramètres mécaniques clés de la paroi cellulaire in vivo et à grande échelle, ce qui a permis les premières observations de la dynamique des parois cellulaires. Ceci a révélé que la paroi cellulaire est plus souple et très variable au niveau des pôles de croissance, et presque stable et plus rigide dans les sites non cultivés. Au cours de l'allongement, il existe une interaction entre la mécanique des parois et la croissance cellulaire, dont le contrôle actif permet l'expansion cellulaire tout en préservant l'intégrité des cellules. De plus, j'ai observé qu'il existe une forte corrélation entre la mécanique des parois cellulaires et la morphologie cellulaire, et des perturbations des propriétés de la paroi affectent directement l'établissement et la maintenance de la forme. Ensemble, mes résultats montrent que la régulation de la paroi est fondamentale dans la détermination de la dynamique cellulaire dans les cellules à parois épaissies. Globalement, cela suggère que l'observation dynamique de la mécanique de surface cellulaire est essentielle pour une compréhension complète des processus multifactoriels et complexes comme la croissance et la morphogenèse

    A Positive Feedback between Growth and Polarity Provides Directional Persistency and Flexibility to the Process of Tip Growth

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    International audiencePolar cell growth is a conserved morphogenetic process needed for survival, mating, and infection [1, 2]. It typically implicates the assembly and spatial stabilization of a cortical polar domain of the active form of a small GTPase of the Rho family, such as Cdc42, which promotes cytoskeleton assembly and secretion needed for local surface expansion [3, 4, 5, 6]. In multiple physiological instances, polarity domains may switch from being spatially unstable, exhibiting a wandering behavior around the cell surface, to being stable at a fixed cellular location [7, 8, 9, 10, 11]. Here, we show that the rate of surface growth may be a key determinant in controlling the spatial stability of active Cdc42 domains. Reducing the growth rate of single rod-shaped fission yeast cells using chemical, genetic, and mechanical means systematically causes polar domains to detach from cell tips and oscillate around the cell surface within minutes. Conversely, an abrupt increase in growth rate improves domain stabilization. A candidate screen identifies vesicular transport along actin cables as an important module mediating this process. Similar behavior observed in distant filamentous fungi suggests that this positive feedback between growth and polarity could represent a basal property of eukaryotic polarization, promoting persistent polar growth as well as growth redirection with respect to the mechanical environment of cells

    Physical Models of Cell Polarity

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    Fission yeast is a pill-shaped unicellular organism, and before dividing it grows by extension at the tips to double the original length. This work consists of mathematical models for how fission yeast controls this growth process. The models presented are either developed in collaboration with experimentalists or using published experimental work on this organism.First, in collaboration with experimentalists Maitreyi Das and Fulvia Verde, we examine the organization of the signaling protein Cdc42, which we implicate as a central part of a control system for polarized growth. Cdc42, a member of the Rho family of proteins, binds to the inner membrane of the cell tips where growth occurs. In collaboration, we find that the fraction of Cdc42 bound to a given cell tip correlates to its growth rate, and that the amount of bound Cdc42 undergoes anti-correlated oscillations between the cell tips. We present a model that describes how Cdc42 and related proteins effect this organization, and shows how the oscillations could function as an exploratory mechanism to help the system overcome a kinetic barrier. Experimental results from our collaborators, such as a loss of correlation in very long cells and a reorganization after disruptive drug treatment, validate the model.Next, using experimental results from literature, we turn to the patterned remodeling of the cell wall. We make a hypothesis that extends the result that Cdc42 marks cell tips for growth from previous work: that Cdc42 marks sites for growth on a microscopic level. A model for the fission yeast cell as an elastic shell being remodeled under turgor pressure at a rate that depends on cortical Cdc42 levels reproduces essential experimental results, namely the ratio of signal width to cell diameter and a linear relation between growth rate and pressure, and gives an estimation of the wall remodeling rate at the cell tips. Since this model predicts that cell diameter depends crucially on the width of a Cdc42 signal, we consider the plausibility of mechanisms for establishing the width of that signal. We find that stronger-than-linear feedback from cell diameter to signal width leads to unstable width regulation, and propose an independent length scale such as from a reaction-diffusion-type mechanism for a cell-diameter-independent Cdc42 signal width. Finally, we describe a mathematical model consisting of Cdc42-signal-dependent cell growth, diffusing Cdc42 growth zones with native width, and an axis-sensing microtubule-based system capable of delivering landmark proteins to the cell tips that bias the diffusion of the growth zones. Parameter dependence of the model is explored, and we show that such a model can give straight, bent, and wide cells, all of which have been observed by experimentalists. We argue that such a model is consistent with the roles of cytoskeleton- and signal-related proteins and known aberrant shapes of mutant cells.As a whole, this work provides mechanistic insight into the system regulating shape and growth in one important model organism

    A focus on yeast mating: From pheromone signaling to cell-cell fusion.

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    Cells live in a chemical environment and are able to orient towards chemical cues. Unicellular haploid fungal cells communicate by secreting pheromones to reproduce sexually. In the yeast models Saccharomyces cerevisiae and Schizosaccharomyces pombe, pheromonal communication activates similar pathways composed of cognate G-protein-coupled receptors and downstream small GTPase Cdc42 and MAP kinase cascades. Local pheromone release and sensing, at a mobile surface polarity patch, underlie spatial gradient interpretation to form pairs between two cells of distinct mating types. Concentration of secretion at the point of cell-cell contact then leads to local cell wall digestion for cell fusion, forming a diploid zygote that prevents further fusion attempts. A number of asymmetries between mating types may promote efficiency of the system. In this review, we present our current knowledge of pheromone signaling in the two model yeasts, with an emphasis on how cells decode the pheromone signal spatially and ultimately fuse together. Though overall pathway architectures are similar in the two species, their large evolutionary distance allows to explore how conceptually similar solutions to a general biological problem can arise from divergent molecular components

    Cell polarization in budding and fission yeasts

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    Polarization is a fundamental cellular property, which is essential for the function of numerous cell types. Over the past three to four decades, research using the best-established yeast systems in cell biological research, Saccharomyces cerevisiae (or budding yeast) and Schizosaccharomyces pombe (or fission yeast), has brought to light fundamental principles governing the establishment and maintenance of a polarized, asymmetric state. These two organisms, though both ascomycetes, are evolutionarily very distant and exhibit distinct shapes and modes of growth. In this review, we compare and contrast the two systems. We first highlight common cell polarization pathways, detailing the contribution of Rho GTPases, the cytoskeleton, membrane trafficking, lipids, and protein scaffolds. We then contrast the major differences between the two organisms, describing their distinct strategies in growth site selection and growth zone dimensions and compartmentalization, which may be the basis for their distinct shape

    How and why cells grow as rods

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    The rod is a ubiquitous shape adopted by walled cells from diverse organisms ranging from bacteria to fungi to plants. Although rod-like shapes are found in cells of vastly different sizes and are constructed by diverse mechanisms, the geometric similarities among these shapes across kingdoms suggest that there are common evolutionary advantages, which may result from simple physical principles in combination with chemical and physiological constraints. Here, we review mechanisms of constructing rod-shaped cells and the bases of different biophysical models of morphogenesis, comparing and contrasting model organisms in different kingdoms. We then speculate on possible advantages of the rod shape, and suggest strategies for elucidating the relative importance of each of these advantages

    Investigating The Regulation of Host Tissue Colonisation by the Rice Blast Fungus Magnaporthe oryzae

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    The filamentous fungus Magnaporthe oryzae is a devastating pathogen of cultivated rice. M. oryzae elaborates a pressurized dome-shaped infection structure, called the appressorium, which physically ruptures the cuticle and gains entry into host tissue. Intracellular invasive hyphae invade neighbouring host cells through plasmodesmata. The Pmk1 MAPK cascade is well known for its roles in regulating the formation and function of the appressorium. Interestingly, ∆pmk1 mutants cannot infect host plant tissue through wounds, suggesting a role in invasive growth. Here, I define biological functions of the Pmk1 MAPK at various stages of the life cycle, by using a controllable version of Pmk1 that is specifically inhibited by a cell-permeable compound without disturbing other wild-type kinases. The Pmk1 MAPK signalling regulates morphogenesis of narrow invasive hyphae traversing the host cell wall, and modulates production of several putative secreted effectors, providing a direct link between the signalling cascade and effector-driven host immune suppression. These results indicate that the Pmk1 pathway is a central regulator of infection-related development necessary for many stages of plant infection including appressorium development, plant penetration, and importantly tissue colonisation. I also report the role of cell cycle progression in the development of plant infection structure. By using two novel conditional mutants that arrest in S and G2 phases, I defined that S-phase progression is crucial for appressorium-mediated plant penetration.The Halpin Trus

    The Dynamical Systems Properties of the HOG Signaling Cascade

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    The High Osmolarity Glycerol (HOG) MAP kinase pathway in the budding yeast Saccharomyces cerevisiae is one of the best characterized model signaling pathways. The pathway processes external signals of increased osmolarity into appropriate physiological responses within the yeast cell. Recent advances in microfluidic technology coupled with quantitative modeling, and techniques from reverse systems engineering have allowed yet further insight into this already well-understood pathway. These new techniques are essential for understanding the dynamical processes at play when cells process external stimuli into biological responses. They are widely applicable to other signaling pathways of interest. Here, we review the recent advances brought by these approaches in the context of understanding the dynamics of the HOG pathway signaling

    La ruta de señalización de MAPK Pmk1p y el mantenimiento de la integridad celular del Schizosaccharomyces Pombe

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    Three cascades of MAPKs have been described about the Schizosaccharomyces pombe such as: the pheromone response with Spk1p as MAP kinase, the stress response in which Sty1p/Spc1p is MAPK, and the maintenance of cell integrity led by Pmk1p/Spm1p. The elimination of any of the kinases of the integrity path causes morphological alterations and multitabicated cells under stress conditions; suggesting a role in ionic homeostasis and cell wall biosynthesis. So, it was proposed to study the role of the MAPK Pmk1p signaling pathway in the maintenance of cell integrity in S.pombe. The organism mainly used was the fission yeast S. pombe and different molecular strains of Escherichia coli were used to carry out the molecular cloning work. Different techniques of molecular cloning, genetic methods, Western Blot and determination of the activity of Pmk1p under stress conditions were used. The most important conclusions were that the "sensors" "Mtl2p and Wsc1p" signal towards Rho1p but they are not "authentic" components of the cascade, and their mutants do not present the Vic phenotype (viable in the presence of immunosuppressant and chloride ion), characteristic of the mutants in the components of the cascade. Mtl2p and Wsc1p do not play an important role in signaling in response to osmotic stress and cell wall damage through the cellular integrity pathway of Pmk1p.Con respecto a la Schizosaccharomyces pombe se han descrito tres cascadas de MAPKs: la de respuesta feromonas, con Spk1p como MAP kinasa; la de respuesta a estrés en la que Sty1p/Spc1p es la MAPK y la de mantenimiento de la integridad celular liderada por Pmk1p/Spm1p. La eliminación de cualquiera de las kinasas de la ruta de integridad provoca alteraciones morfológicas y células multitabicadas en condiciones de estrés; lo que sugiere una función en homeostasis iónica y en la biosíntesis de la pared celular por lo que se propuso estudiar el papel de la ruta de señalización de MAPK Pmk1p en el mantenimiento de la integridad celular en S.pombe. El organismo mayoritariamente utilizado fue la levadura de fisión S. pombe y para realizar los trabajos de clonación molecular se utilizaron también diferentes estirpes de Escherichia coli. Se emplearon diversas técnicas de clonación molecular, métodos genéticos, Western Blot y determinación de la actividad de Pmk1p bajo condiciones de estrés. Las conclusiones más importantes fueron: que los “sensores” “Mtl2p y Wsc1p” señalizan hacia Rho1p pero no son componentes “auténticos” de la cascada, y sus mutantes no presentan el fenotipo vic (viable en presencia de inmunosupresor y de iones cloruro), característico de los mutantes en los componentes de la cascada. Mtl2p y Wsc1p no desempeñan un papel importante en la señalización en respuesta a estrés osmótico y daño en la pared celular a través de la ruta de integridad celular de Pmk1p
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