Functional characterisation of Arabidopsis SPL7 conserved protein domains suggests novel regulatory mechanisms in the Cu deficiency response

BACKGROUND: The Arabidopsis SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE (SPL) transcription factor SPL7 reprograms cellular gene expression to adapt plant growth and cellular metabolism to copper (Cu) limited culture conditions. Plant cells require Cu to maintain essential processes, such as photosynthesis, scavenging reactive oxygen species, cell wall lignification and hormone sensing. More specifically, SPL7 activity promotes a high-affinity Cu-uptake system and optimizes Cu (re-)distribution to essential Cu-proteins by means of specific miRNAs targeting mRNA transcripts for those dispensable. However, the functional mechanism underlying SPL7 activation is still to be elucidated. As SPL7 transcript levels are largely non-responsive to Cu availability, post-translational modification seems an obvious possibility. Previously, it was reported that the SPL7 SBP domain does not bind to DNA in vitro in the presence of Cu ions and that SPL7 interacts with a kin17 domain protein to raise SPL7-target gene expression upon Cu deprivation. Here we report how additional conserved SPL7 protein domains may contribute to the Cu deficiency response in Arabidopsis. RESULTS: Cytological and biochemical approaches confirmed an operative transmembrane domain (TMD) and uncovered a dual localisation of SPL7 between the nucleus and an endomembrane system, most likely the endoplasmic reticulum (ER). This new perspective unveiled a possible link between Cu deficit and ER stress, a metabolic dysfunction found capable of inducing SPL7 targets in an SPL7-dependent manner. Moreover, in vivo protein-protein interaction assays revealed that SPL7 is able to homodimerize, probably mediated by the IRPGC domain. These observations, in combination with the constitutive activation of SPL7 targets, when ectopically expressing the N-terminal part of SPL7 including the SBP domain, shed some light on the mechanisms governing SPL7 function. CONCLUSIONS: Here, we propose a revised model of SPL7 activation and regulation. According to our results, SPL7 would be initially located to endomembranes and activated during ER stress as a result of Cu deficiency. Furthermore, we added the SPL7 dimerization in the presence of Cu ions as an additional regulatory mechanism to modulate the Cu deficiency response. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12870-014-0231-5) contains supplementary material, which is available to authorized users

A Conserved KIN17 Curved DNA-Binding Domain Protein Assembles with SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE7 to Adapt Arabidopsis Growth and Development to Limiting Copper Availability

Proper copper (Cu) homeostasis is required by living organisms to maintain essential cellular functions. In the model plant Arabidopsis (Arabidopsis thaliana), the SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE7 (SPL7) transcription factor participates in reprogramming global gene expression during Cu insufficiency in order to improve the metal uptake and prioritize its distribution to Cu proteins of major importance. As a consequence, spl7 null mutants show morphological and physiological disorders during Cu-limited growth, resulting in lower fresh weight, reduced root elongation, and chlorosis. On the other hand, the Arabidopsis KIN17 homolog belongs to a well-conserved family of essential eukaryotic nuclear proteins known to be stress activated and involved in DNA and possibly RNA metabolism in mammals. In the study presented here, we uncovered that Arabidopsis KIN17 participates in promoting the Cu deficiency response by means of a direct interaction with SPL7. Moreover, the double mutant kin17-1 spl7-2 displays an enhanced Cu-dependent phenotype involving growth arrest, oxidative stress, floral bud abortion, and pollen inviability. Taken together, the data presented here provide evidence for SPL7 and KIN17 protein interaction as a point of convergence in response to both Cu deficiency and oxidative stress

The Copper-microRNA Pathway Is Integrated with Developmental and Environmental Stress Responses in Arabidopsis thaliana

As an essential nutrient, copper (Cu) scarcity causes a decrease in agricultural production. Cu deficiency responses include the induction of several microRNAs, known as Cu-miRNAs, which are responsible for degrading mRNAs from abundant and dispensable cuproproteins to economize copper when scarce. Cu-miRNAs, such as miR398 and miR408 are conserved, as well as the signal transduction pathway to induce them under Cu deficiency. The Arabidopsis thaliana SQUAMOSA-PROMOTER BINDING PROTEIN-LIKE (SPL) family member SPL7 binds to the cis-regulatory motifs present in the promoter regions of genes expressed under Cu deficiency, including Cu-miRNAs. The expression of several other SPL transcription factor family members is regulated by miR156. This regulatory miR156-SPL module plays a crucial role in developmental phase transitions while integrating internal and external cues. Here, we show that Cu deficiency also affects miR156 expression and that SPL3 overexpressing plants, resistant to miR156 regulation, show a severe decrease in SPL7-mediated Cu deficiency responses. These include the expression of Cu-miRNAs and their targets and is probably due to competition between SPL7 and miR156-regulated SPL3 in binding to cis-regulatory elements in Cu-miRNA promoters. Thus, the conserved SPL7-mediated Cu-miRNA pathway could generally be affected by the miR156-SPL module, thereby underscoring the integration of the Cu-miRNA pathway with developmental and environmental stress responses in Arabidopsis thaliana.</i

Equal latency contours and auditory weighting functions for the harbour porpoise (Phocoena phocoena)

This work was supported by The Netherlands Ministry of Infrastructure and the Environment [grant number 4500182046], and by matched funding from The Netherlands Ministry of Defence (administered by TNO) and the UK Natural Environment Research Council [to P.J.W.].Loudness perception by human infants and animals can be studied under the assumption that sounds of equal loudness elicit equal reaction times (RTs). Simple RTs of a harbour porpoise to narrowband frequency-modulated signals were measured using a behavioural method and an RT sensor based on infrared light. Equal latency contours, which connect equal RTs across frequencies, for reference values of 150-200 ms (10 ms intervals) were derived from median RTs to 1 s signals with sound pressure levels (SPLs) of 59-168 dB re. 1 μPa and centre frequencies of 0.5, 1, 2, 4, 16, 31.5, 63, 80 and 125 kHz. The higher the signal level was above the hearing threshold of the harbour porpoise, the quicker the animal responded to the stimulus (median RT 98-522 ms). Equal latency contours roughly paralleled the hearing threshold at relatively low sensation levels (higher RTs). The difference in shape between the hearing threshold and the equal latency contours was more pronounced at higher levels (lower RTs); a flattening of the contours occurred for frequencies below 63 kHz. Relationships of the equal latency contour levels with the hearing threshold were used to create smoothed functions assumed to be representative of equal loudness contours. Auditory weighting functions were derived from these smoothed functions that may be used to predict perceived levels and correlated noise effects in the harbour porpoise, at least until actual equal loudness contours become available.Publisher PDFPeer reviewe

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Captivated by thought:"Sticky" thinking leaves traces of perceptual decoupling in task-evoked pupil size

Throughout the day, we may sometimes catch ourselves in patterns of thought that we experience as rigid and difficult to disengage from. Such "sticky" thinking can be highly disruptive to ongoing tasks, and when it turns into rumination constitutes a vulnerability for mental disorders such as depression and anxiety. The main goal of the present study was to explore the stickiness dimension of thought, by investigating how stickiness is reflected in task performance and pupil size. To measure spontaneous thought processes, we asked participants to perform a sustained attention to response task (SART), in which we embedded the participant's concerns to potentially increase the probability of observing sticky thinking. The results indicated that sticky thinking was most frequently experienced when participants were disengaged from the task. Such episodes of sticky thought could be discriminated from neutral and non-sticky thought by an increase in errors on infrequent no-go trials. Furthermore, we found that sticky thought was associated with smaller pupil responses during correct responding. These results demonstrate that participants can report on the stickiness of their thought, and that stickiness can be investigated using pupillometry. In addition, the results suggest that sticky thought may limit attention and exertion of cognitive control to the task

Rotaxane-Functionalized Dyes for Charge-Rectification in<i> p</i>-Type Photoelectrochemical Devices

A supramolecular photovoltaic strategy is applied to enhance power conversion efficiencies (PCE) of photoelectrochemical devices by suppressing electron–hole recombination after photoinduced electron transfer (PET). Here, the author exploit supramolecular localization of the redox mediator—in close proximity to the dye—through a rotaxane topology, reducing electron–hole recombination in p-type dye-sensitized solar cells (p-DSSCs). Dye PRotaxane features 1,5-dioxynaphthalene recognition sites (DNP-arms) with a mechanically-interlocked macrocyclic redox mediator naphthalene diimide macrocycle (3-NDI-ring), stoppering synthetically via click chemistry. The control molecule PStopper has stoppered DNP-arms, preventing rotaxane formation with the 3-NDI-ring. Transient absorption and time-resolved fluorescence spectroscopy studies show ultrafast (211 ± 7 fs and 2.92 ± 0.05 ps) PET from the dye-moiety of PRotaxane to its mechanically interlocked 3-NDI-ring-acceptor, slowing down the electron–hole recombination on NiO surfaces compared to the analogue. p-DSSCs employing PRotaxane (PCE = 0.07%) demonstrate a 30% PCE increase compared to PStopper (PCE = 0.05%) devices, combining enhancements in both open-circuit voltages (VOC = 0.43 vs 0.36 V) and short-circuit photocurrent density (JSC = −0.39 vs −0.34 mA cm−2). Electrochemical impedance spectroscopy shows that PRotaxane devices exhibit hole lifetimes (τh) approaching 1 s, a 16-fold improvement compared to traditional I−/I3−-based systems (τh = 50 ms), demonstrating the benefits obtained upon nanoengineering of interfacial dye-regeneration at the photocathode.</p

Issues of belonging, pedagogy and learning in doctoral study at a distance

In this paper we present a case study of doctoral study at a distance, and we explore issues of belonging, pedagogy and learning as part of that process. As a team of one doctoral researcher and three supervisors, we critically reflect on the place of belonging in the context of doctoral study by distance. In this case study, the importance of belonging was heightened due to a high-risk and highly volatile context in which the doctoral researcher lived, the Kurdistan Region of Iraq. We further explore the elements that developed a sense of belonging, aided by a range of digital technologies. Our findings suggest that the place of belonging in learning needs further examination in higher education contexts, especially when universities are keen to increase distance enrolments

21st century skills, problem based learning and the university of the future

In this paper we focus on the ‘university of the future’, and in particular we aim to explore what the role and relevance of PBL might be in this imagined university of the future. We explore the potential of PBL in the development of 21st century skills in a higher education environment that is on the brink of profound changes, and that consequently requires a continuously adaptive approach to education renewal at all levels. Of course many of these changes are already underway. We appear to be in the midst of profound disruptions to higher education, not in the least due to fast changing technologies and the possibilities they afford. The internet and the World Wide Web have had huge impacts, which in turn have influenced the social fabric of our lives through the growing ubiquity of social media and mobile media tools. In education in general, and in higher education in particular, these changes have ushered in an age characterised by a rapidly increasing evolution of online learning with integration of online, hybrid, and collaborative learning, and most recently, phenomena such as Massive Open Online Courses (MOOCs), the rise of big data analytics driving learning, and personalised learning. Each of these developments have the potential to cause major disruptions in the way we operate in higher education. It is important to recognise and respect that these changes are here to stay, some evolutionary, some revolutionary. So we need to respond in adaptive and agile ways, and importantly, with imagination and creativity. In this paper, we explore the potential of PBL to address some of the unknown and uncertain challenges of the 21st century