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De Bortoli, Sara (2017) Characterization of ion channels in chloroplasts and mitochondria of land plants. [Ph.D. thesis]

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Abstract (english)

Chloroplasts and mitochondria play essential roles in the plant physiology and are emerging as important players in intracellular Ca2+ signaling. A major role in the context of organellar ion homeostasis is played by ion channels. In fact, they are responsible for the regulation of ion distribution between compartments and they are proposed to guarantee Ca2+ signaling, proper osmotic potential, optimal pH for enzymatic activities and electron transport chains’ function. In plants, only a few of channel families have been found to localize to chloroplast/mitochondria and most of them have not been fully characterized. In addition, the molecular mechanisms by which chloroplasts and mitochondria accumulate and release Ca2+ are still far from being clarified. The present work aimed to characterize possible players in organellar Ca2+ flux-mediating systems in plants, namely the Ionotropic Glutamate Receptors (GLRs, non-selective cation channels) and Mitochondrial Calcium Uniporters (MCUs), combining reverse genetics, biochemistry and in vivo localization studies.
The research here presented allowed to deepen the study on land plant GLRs, to characterize two novel organellar channels of A. thaliana, i. e. AtGLR3.5 and AtMCU, and to lay the ground for the characterization of the Physcomitrella patens GLRS, PpGLRs, from different points of view. Results revealed additional variability of GLRs among photosynthetic organisms and support the role of GLRs in the regulation of various organellar processes such as photoprotective mechanisms, senescence and mitochondrial structure integrity maintenance. Importantly, the characterization of AtGLR3.5 provided the molecular identification of the first cation channel in plant mitochondria and the characterization of AtMCU1 and MICU allowed for the first time the proposal of a model for the existence and regulation of Ca2+ fluxes in plant mitochondria via MCU complex. The present work therefore contributed to add new knowledge to the field of the regulation of ion homeostasis, especially Ca2+ homeostasis, in mitochondria and chloroplasts.

Abstract (italian)

Cloroplasti e mitocondri hanno un ruolo fondamentale nella fisiologia vegetale e stanno emergendo come importanti attori nel signaling del Ca2+ intracellulare. Un ruolo fondamentale nel contesto dell’omeostasi ionica organellare è giocato dai canali ionici. Infatti essi sono responsabili della regolazione della distribuzione ionica fra compartimenti ed è stato proposto che contribuiscano a garantire il signaling del Ca2+, l’appropriato potenziale osmotico, il pH ottimale per le attività enzimatiche e il funzionamento delle catene di trasporto elettronico. Nelle piante, solo poche famiglie di canali sono state identificate in cloroplasti/mitocondri e la maggior parte di loro non è stata pienamente caratterizzata. Inoltre, il meccanismo molecolare attraverso cui cloroplasti e mitocondri accumulano e rilasciano Ca2+ è ancora lontano dall’essere chiarito. Lo scopo del presente lavoro è stato quello di caratterizzare possibili attori coinvolti nel nella mediazione dei flussi di Ca2+ organellari, in particolare i recettori ionotropici del glutammato (GLRs) e gli uniporti mitocondriali del calcio (MCU), combinando tecniche di genetica inversa, biochimica e studi di localizzazione in vivo.
La ricerca qui presentata ha permesso di approfondire lo studio sui GLR vegetali, di caratterizzare due nuovi canali organellari di A. thaliana, AtGLR3.5 e AtMCU, e di gettare le fondamenta per la caratterizzazione dei GLR di Physcomitrella patens, PpGLR, da diversi punti di vista. I risultati del lavoro hanno rivelato ulteriore variabilità dei GLR fra gli organismi fotosintetici e hanno mostrato come i GLR siano coinvolti nella regolazione di diversi processi organellari come i meccanismi di fotoprotezione, la senescenza e il mantenimento dell’integrità strutturale mitocondriale. La caratterizzazione di AtGLR3.5 ha permesso l’identificazione molecolare del primo canale cationico mitocondriale vegetale e la caratterizzazione di AtMCU1 ha permesso per la prima volta di proporre un modello che preveda l’esistenza e la regolazione dei flussi di Ca2+ nei mitocondri vegetali per mezzo del complesso MCU. Il lavoro qui presentato ha pertanto contribuito ad aggiungere nuova conoscenza al campo della regolazione dell’omeostasi ionica, specialmente l’omeostasi del Ca2+, nei mitocondri e nei cloroplasti.

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EPrint type:Ph.D. thesis
Tutor:Szabo', Ildiko'
Supervisor:Teardo, Enrico
Ph.D. course:Ciclo 29 > Corsi 29 > BIOSCIENZE E BIOTECNOLOGIE
Data di deposito della tesi:31 January 2017
Anno di Pubblicazione:31 January 2017
Key Words:ionotropic glutamate receptors mitochondrial calcium uniporters recettore ionotropico del glutammato uniporto mitocondriale del calcio calcium signaling homeostasis omeostasi ion ioni arabidopsis physcomitrella
Settori scientifico-disciplinari MIUR:Area 05 - Scienze biologiche > BIO/10 Biochimica
Struttura di riferimento:Dipartimenti > Dipartimento di Biologia
Codice ID:10282
Depositato il:17 Nov 2017 09:23
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