FerriTag is a new genetically-encoded inducible tag for correlative light-electron microscopy

A current challenge is to develop tags to precisely visualize proteins in cells by light and electron microscopy. Here, we introduce FerriTag, a genetically-encoded chemically-inducible tag for correlative light-electron microscopy. FerriTag is a fluorescent recombinant electron-dense ferritin particle that can be attached to a protein-of-interest using rapamycin-induced heterodimerization. We demonstrate the utility of FerriTag for correlative light-electron microscopy by labeling proteins associated with various intracellular structures including mitochondria, plasma membrane, and clathrin-coated pits and vesicles. FerriTagging has a good signal-to-noise ratio and a labeling resolution of approximately 10 nm. We demonstrate how FerriTagging allows nanoscale mapping of protein location relative to a subcellular structure, and use it to detail the distribution and conformation of huntingtin-interacting protein 1 related (HIP1R) in and around clathrin-coated pits

Clipping

We are living in a new era of haunting, one in which (as the essays published here suggest) we find ourselves engaging with the 'ethics of the spectral text', 'spectral and textual haunting', and 'ghostly narrative' (as distinct from narrative about the ghostly). Such phrases themselves tell a story or at least evoke a disorder that is in keeping with this new era. They are new kinds of phrases: they seek a place, a haunt, in which spectrality cohabits with writing, text and narrative, as well as with the question of ethics.You have to move very fast. Everything is just clipping by. That's what is at issue in this new era in which hauntology replaces ontology and increasing spectralization is the disorder of the day

Reproductive decisions in the lesser black-backed gull larus fuscus and their effects on reproductive success

The effect of several fundamental reproductive 'decisions' upon reproductive success were examined over a three year period at a large, inland gullery in the Pennines. Variations in reproductive parameters in relation to timing of breeding and reproductive success were compared among years. Determinants of the degree of hatching asynchrony were identified. Eggs from two years were taken for yolk lipid analysis, using gas chromatography. Variation in micronutrient content of eggs within clutches and between years was assessed in relation to egg size and yolk size, in order to examine resource allocation decisions of individuals. Timing of breeding of individuals was experimentally manipulated through the exchange of whole clutches of eggs between early and late laying birds, whilst controlling for variation in clutch size and egg-size, in order to assess whether the seasonal decrease in reproductive success was best explained by a decrease in food supply or differences in quality among parents. I experimentally manipulated the within-brood mass hierarchy of gulls, whilst controlling for variation in both chick quality and parental quality, in order to assess the effect of hatching asynchrony per se on chick growth and survival, and whether parents optimized the degree of hatching asynchrony with respect to the prevailing food supply. Brood size was experimentally reduced in order to assess the costs and benefits of the production of supernumary young. This was acheived by comparison of chick growth, feather development and chick survival of unmanipulated three-chick broods with broods where either the a-chick or the c- chick had been removed. I present a general discussion of the results within the context of life-history theory and a model for the evolution of hatching asynchrony in the lesser black-backed gul

Clipping

We are living in a new era of haunting, one in which (as the essays published here suggest) we find ourselves engaging with the 'ethics of the spectral text', 'spectral and textual haunting', and 'ghostly narrative' (as distinct from narrative about the ghostly). Such phrases themselves tell a story or at least evoke a disorder that is in keeping with this new era. They are new kinds of phrases: they seek a place, a haunt, in which spectrality cohabits with writing, text and narrative, as well as with the question of ethics.You have to move very fast. Everything is just clipping by. That's what is at issue in this new era in which hauntology replaces ontology and increasing spectralization is the disorder of the day

Theorizing theory or the future of theory

Interview conducted by Petar Pend

The stories we tell: uncanny encounters in Mr Straw’s house

During my first visit to Mr Straw’s House, a National Trust Property in the North of England, I was intrigued by the discrepancies between the narrative framework provided by the National Trust – its exclusions, silences and invisibilities – and the far more complex stories the house seemed to tantalisingly hint at. As a scholar I am drawn to certain sites and affectively engage with them and yet I usually keep silent about my investment which informs not only my interest but also how I read these heritage sites. My aim here is not primarily to interrogate my own investment, but to ask how productive it is, what it enables me to see and to describe and where its limits are. This case study explores a particular tourist attraction from the perspective of storytelling and asks what narratives can be constructed around, and generated through, the spatial-emotional dimensions of this heritage site. I am interested in the hold sites have over people, why and how they provoke imaginative and empathic investment that generates a network of stories and triggers processes of unravelling which have the potential to transform silences and unmetabolised affect into empathy and emotional thought

Congressing kinetochores progressively load Ska complexes to prevent force-dependent detachment

Kinetochores mediate chromosome congression by either sliding along the lattice of spindle microtubules or forming end-on attachments to their depolymerizing plus-ends. By following the fates of individual kinetochores as they congress in live cells, we reveal that the Ska complex is required for a distinct substep of the depolymerization-coupled pulling mechanism. Ska depletion increases the frequency of naturally occurring, force-dependent P kinetochore detachment events, while being dispensable for the initial biorientation and movement of chromosomes. In unperturbed cells, these release events are followed by reattachment and successful congression, whereas in Ska-depleted cells, detached kinetochores remain in a futile reattachment/detachment cycle that prevents congression. We further find that Ska is progressively loaded onto bioriented kinetochore pairs as they congress. We thus propose a model in which kinetochores mature through Ska complex recruitment and that this is required for improved load-bearing capacity and silencing of the spindle assembly checkpoint

Tumor protein D54 defines a new class of intracellular transport vesicles

Transport of proteins and lipids from one membrane compartment to another is via intracellular vesicles. We investigated the function of tumor protein D54 (TPD54/TPD52L2) and found that TPD54 was involved in multiple membrane trafficking pathways: anterograde traffic, recycling, and Golgi integrity. To understand how TPD54 controls these diverse functions, we used an inducible method to reroute TPD54 to mitochondria. Surprisingly, this manipulation resulted in the capture of many small vesicles (30 nm diameter) at the mitochondrial surface. Super-resolution imaging confirmed the presence of similarly sized TPD54-positive structures under normal conditions. It appears that TPD54 defines a new class of transport vesicle, which we term intracellular nanovesicles (INVs). INVs meet three criteria for functionality. They contain specific cargo, they have certain R-SNAREs for fusion, and they are endowed with a variety of Rab GTPases (16 out of 43 tested). The molecular heterogeneity of INVs and the diverse functions of TPD54 suggest that INVs have various membrane origins and a number of destinations. We propose that INVs are a generic class of transport vesicle that transfer cargo between these varied locations

The Digital Journey: Reflected Learnings and Emerging Challenges

This paper provides a literature-based examination of the four stages of marketing’s digital journey to date and the key milestones or points of interest along the way. In reflecting on the path taken, intriguing insights emerge which pose ongoing challenges for businesses and for the marketing discipline. Arguably failure to heed the resulting warnings will impede marketing’s ability to harness the benefits of digital and to effectively counteract the associated negative consequences of a digitally-enabled marketplace. The implications of marketing’s digital journey are explored here along with their most pressing consequences. This is the peer reviewed version of the following article: Quinton, S & Simkin, L 2016, 'The Digital Journey: Reflected Learnings and Emerging Challenges' International Journal of Management Reviews, vol In press. DOI: 10.1111/ijmr.12104, which has been published in final form at https://dx.doi.org/10.1111/ijmr.12104 This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving