Senior Recital: Emily Breeze, mezzo-soprano

https://digitalwindow.vassar.edu/musi_senior/1004/thumbnail.jp

The Passage of H₂O₂ from Chloroplasts to Their Associated Nucleus during Retrograde Signalling: Reflections on the Role of the Nuclear Envelope.

The response of chloroplasts to adverse environmental cues, principally increases in light intensity, stimulates chloroplast-to-nucleus retrograde signalling, which leads to the induction of immediate protective responses and longer-term acclimation. Hydrogen peroxide (H2O2), generated during photosynthesis, is proposed to both initiate and transduce a retrograde signal in response to photoinhibitory light intensities. Signalling specificity achieved by chloroplast-sourced H2O2 for signal transduction may be dependent upon the oft-observed close association of a proportion of these organelles with the nucleus. In this review, we consider more precisely the nature of the close association between a chloroplast appressed to the nucleus and the requirement for H2O2 to cross both the double membranes of the chloroplast and nuclear envelopes. Of particular relevance is that the endoplasmic reticulum (ER) has close physical contact with chloroplasts and is contiguous with the nuclear envelope. Therefore, the perinuclear space, which transducing H2O2 molecules would have to cross, may have an oxidising environment the same as the ER lumen. Based on studies in animal cells, the ER lumen may be a significant source of H2O2 in plant cells arising from the oxidative folding of proteins. If this is the case, then there is potential for the ER lumen/perinuclear space to be an important location to modify chloroplast-to-nucleus H2O2 signal transduction and thereby introduce modulation of it by additional different environmental cues. These would include for example, heat stress and pathogen infection, which induce the unfolded protein response characterised by an increased H2O2 level in the ER lumen

Arabidopsis Lunapark proteins are involved in ER cisternae formation

(1) The plant endoplasmic reticulum (ER) is crucial to the maintenance of cellular homeostasis. The ER consists of a dynamic and continuously remodelling network of tubules and cisternae. Several conserved membrane proteins have been implicated in formation and maintenance of the ER network in plants, such as RHD3 and the reticulon proteins. Despite the recent work in mammalian and yeast cells, the detailed molecular mechanisms of ER network organisation in plants still remain largely unknown. Recently novel ER network-shaping proteins called Lunapark (LNP) have been identified in yeast and mammalian cells. (2) Here we identify two arabidopsis LNP homologues and investigate their subcellular localisation via confocal microscopy and potential function in shaping the ER network using protein-protein interaction assays and mutant analysis. (3) We show that AtLNP1 overexpression in tobacco leaf epidermal cells mainly labels cisternae in the ER network whereas AtLNP2 labels the whole ER. Overexpression of LNP proteins results in an increased abundance of ER cisternae and lnp1 and lnp1lnp2 amiRNA lines display a reduction in cisternae and larger polygonal areas. (4) Thus, we hypothesize that AtLNP1 and AtLNP2 are involved in determining the network morphology of the plant ER, possibly by regulating the formation of ER cisternae

The plant endoplasmic reticulum is both receptive and responsive to pathogen effectors

The endoplasmic reticulum (ER) is the entry point to the secretory pathway and, as such, is critical for adaptive responses to biotic stress, when the demand for de novo synthesis of immunity-related proteins and signalling components increases significantly. Comprised of a network of interconnected tubules and cisternae, the architecture of the ER is highly pleomorphic and dynamic, rapidly remodelling to meet new cellular requirements. During infection with the hemi-biotrophic phytopathogen, Pseudomonas syringae pv. tomato DC3000, the ER in cells immediately adjacent to established bacterial colonies condenses into ‘knot-like’ structures, reminiscent of fenestrated sheets. Based on known temporal dynamics of pathogen effector delivery and initial bacterial multiplication, the timing of these observed morphological changes is rapid and independent of classical elicitor activation of pathogen-triggered immunity. To further investigate a role for ER reconfiguration in suppression of plant immunity we identified a conserved C-terminal tail-anchor domain in a set of pathogen effectors known to localize to the ER and used this protein topology in an in silico screen to identify putative ER-localised effectors within the effectorome of the oomycete Phytophthora infestans. Subsequent characterization of a subset of 15 candidate tail- anchored P. infestans effectors revealed that 11 localised to the ER and/or Golgi. Notably, transient expression of an ER-localised effector from the closely related oomycete, Plasmopara halstedii, reconfigured the ER network, revealing intimate association of labelled ER with perinuclear chloroplasts and clusters of chloroplasts, potentially facilitating retrograde signalling during plant defence