Scribble scrambles parathyroid hormone receptor interactions to regulate phosphate and vitamin D homeostasis.

G protein-coupled receptors, including PTHR, are pivotal for controlling metabolic processes ranging from serum phosphate and vitamin D levels to glucose uptake, and cytoplasmic interactors may modulate their signaling, trafficking, and function. We now show that direct interaction with Scribble, a cell polarity-regulating adaptor protein, modulates PTHR activity. Scribble is a crucial regulator for establishing and developing tissue architecture, and its dysregulation is involved in various disease conditions, including tumor expansion and viral infections. Scribble co-localizes with PTHR at basal and lateral surfaces in polarized cells. Using X-ray crystallography, we show that colocalization is mediated by engaging a short sequence motif at the PTHR C-terminus using Scribble PDZ1 and PDZ3 domain, with binding affinities of 31.7 and 13.4 µM, respectively. Since PTHR controls metabolic functions by actions on renal proximal tubules, we engineered mice to selectively knockout Scribble in proximal tubules. The loss of Scribble impacted serum phosphate and vitamin D levels and caused significant plasma phosphate elevation and increased aggregate vitamin D3 levels, whereas blood glucose levels remained unchanged. Collectively these results identify Scribble as a vital regulator of PTHR-mediated signaling and function. Our findings reveal an unexpected link between renal metabolism and cell polarity signaling.

Structural analysis of human papillomavirus E6 interactions with Scribble PDZ domains.

The cell polarity regulator Scribble has been shown to be a critical regulator of the establishment and development of tissue architecture, and its dysregulation promotes or suppresses tumour development in a context-dependent manner. Scribble activity is subverted by numerous viruses. This includes human papillomaviruses (HPVs), who target Scribble via the E6 protein. Binding of E6 from high-risk HPV strains to Scribble via a C-terminal PDZ-binding motif leads to Scribble degradation in vivo. However, the precise molecular basis for Scribble-E6 interactions remains to be defined. We now show that Scribble PDZ1 and PDZ3 are the major interactors of HPV E6 from multiple high-risk strains, with each E6 protein displaying a unique interaction profile. We then determined crystal structures of Scribble PDZ1 and PDZ3 domains in complex with the PDZ-binding motif (PBM) motifs of E6 from HPV strains 16, 18 and 66. Our findings reveal distinct interaction patterns for each E6 PBM motif from a given HPV strain, suggesting that a complex molecular interplay exists that underpins the overt Scribble-HPV E6 interaction and controls E6 carcinogenic potential.

Persistent retroperitoneal haematoma from undiagnosed renal cell carcinoma in a young trauma patient.

A man in his 20s presents to the emergency department after a water skiing accident and was diagnosed with a grade 3 left renal laceration. He subsequently required cystoscopic insertion of a ureteric stent after failing a trial of conservative management. Over the next 9 months, he re-presented to the hospital twice with increasing flank pain and fevers. Subsequent imaging demonstrated interval progression of the retroperitoneal haematoma with a suspicious calcified lower pole lesion which was biopsied subsequently and revealed malignant tissue. External compression of the kidney by this large haematoma was also thought to be contributing to a state of Page kidney. The patient underwent definitive management with an open left-sided radical nephrectomy which confirmed type 2 papillary renal cell carcinoma. The patient is now normotensive and back to his baseline function. He will undergo surveillance CT imaging and be referred to familial genetic services.

A global horizon scan of issues impacting marine and coastal biodiversity conservation.

The biodiversity of marine and coastal habitats is experiencing unprecedented change. While there are well-known drivers of these changes, such as overexploitation, climate change and pollution, there are also relatively unknown emerging issues that are poorly understood or recognized that have potentially positive or negative impacts on marine and coastal ecosystems. In this inaugural Marine and Coastal Horizon Scan, we brought together 30 scientists, policymakers and practitioners with transdisciplinary expertise in marine and coastal systems to identify new issues that are likely to have a significant impact on the functioning and conservation of marine and coastal biodiversity over the next 5-10 years. Based on a modified Delphi voting process, the final 15 issues presented were distilled from a list of 75 submitted by participants at the start of the process. These issues are grouped into three categories: ecosystem impacts, for example the impact of wildfires and the effect of poleward migration on equatorial biodiversity; resource exploitation, including an increase in the trade of fish swim bladders and increased exploitation of marine collagens; and new technologies, such as soft robotics and new biodegradable products. Our early identification of these issues and their potential impacts on marine and coastal biodiversity will support scientists, conservationists, resource managers and policymakers to address the challenges facing marine ecosystems.

Depth and habitat are important drivers of abundance for predatory reef fish off Pemba Island, Tanzania.

Coral reefs across the world face significant threats from fishing and climate change, which tends to be most acute in shallower waters. This is the case off Pemba Island, Tanzania, yet the effects of these anthropogenic stressors on the distribution and abundance of economically and ecologically important predatory reef fish, including how they vary with depth and habitat type, is poorly understood. Thus, we deployed 79 baited remote underwater videos (BRUVs) in variable water depths and habitats off Pemba Island, and modeled the effects of depth and habitat on abundance of predatory reef fish. Predatory reef fish types/taxa were significantly predicted by depth and habitat types. Habitats in relatively deeper waters and dominated by hard and soft corals hosted high species richness and abundance of predatory reef fish types/taxa compared to mixed sandy and rubble habitats. The findings add to the growing evidence that deep waters around coral reefs are important habitats for predatory reef fish. Thus, careful management, through effective area and species protection measures, is needed to prevent further depletion of predatory reef-associated fish populations and to conserve this biologically important area.

Protection outcomes for fish trophic groups across a range of management regimes.

Understanding how Marine Protected Areas (MPAs) improve conservation outcomes across anthropogenic pressures can improve the benefits derived from them. Effects of protection for coral reefs in the western and central Indian Ocean were assessed using size-spectra analysis of fish and the relationships of trophic group biomass with human population density. Length-spectra relationships quantifying the relative abundance of small and large fish (slope) and overall productivity of the system (intercept) showed inconsistent patterns with MPA protection. The results suggest that both the slopes and intercepts were significantly higher in highly and well-protected MPAs. This indicates that effective MPAs are more productive and support higher abundances of smaller fish, relative to moderately protected MPAs. Trophic group biomass spanning piscivores and herbivores, decreased with increasing human density implying restoration of fish functional structure is needed. This would require addressing fisher needs and supporting effective MPA management to secure ecosystem benefits for coastal communities.

Crystallographic Studies of PDZ Domain-Peptide Interactions of the Scribble Polarity Module.

The determination of high-resolution crystal structures of cell polarity regulatory proteins bound to their functional interactors has proven to be invaluable for deciphering the underlying molecular mechanisms. Here we describe methods to identify suitable complexes of cell polarity protein domains bound to interacting ligands with subsequent preparation of such complexes for X-ray crystallographic analysis.

Metal pollution as a potential threat to shell strength and survival in marine bivalves.

Marine bivalve molluscs, such as scallops, mussels and oysters, are crucial components of coastal ecosystems, providing a range of ecosystem services, including a quarter of the world's seafood. Unfortunately, coastal marine areas often suffer from high levels of metals due to dumping and disturbance of contaminated material. We established that increased levels of metal pollution (zinc, copper and lead) in sediments near the Isle of Man, resulting from historical mining, strongly correlated with significant weakening of shell strength in king scallops, Pecten maximus. This weakness increased mortality during fishing and left individuals more exposed to predation. Comparative structural analysis revealed that shells from the contaminated area were thinner and exhibited a pronounced mineralisation disruption parallel to the shell surface within the foliated region of both the top and bottom valves. Our data suggest that these disruptions caused reduced fracture strength and hence increased mortality, even at subcritical contamination levels with respect to current international standards. This hitherto unreported effect is important since such non-apical responses rarely feed into environmental quality assessments, despite potentially significant implications for the survival of organisms exposed to contaminants. Hence our findings highlight the impact of metal pollution on shell mineralisation in bivalves and urge a reappraisal of currently accepted critical contamination levels.

Structural analysis of phosphorylation-associated interactions of human MCC with Scribble PDZ domains.

Scribble is a crucial adaptor protein that plays a pivotal role during establishment and control of cell polarity, impacting many physiological processes ranging from cell migration to immunity and organization of tissue architecture. Scribble harbours a leucine-rich repeat domain and four PDZ domains that mediate most of Scribble's interactions with other proteins. It has become increasingly clear that post-translational modifications substantially impact Scribble-ligand interactions, with phosphorylation being a major modulator of binding to Scribble. To better understand how Scribble PDZ domains direct cell polarity signalling and how phosphorylation impacts this process, we investigated human Scribble interactions with MCC (Mutated in Colorectal Cancer). We systematically evaluated the ability of all four individual Scribble PDZ domains to bind the PDZ-binding motif (PBM) of MCC as well as MCC phosphorylated at the -1 Ser position. We show that Scribble PDZ1 and PDZ3 are the major interactors with MCC, and that modifications to Ser at the -1 position in the MCC PBM only has a minor effect on binding to Scribble PDZ domains. We then examined the structural basis for these observations by determining the crystal structures of Scribble PDZ1 domain bound to both the unphosphorylated MCC PBM as well as phosphorylated MCC. Our structures indicated that phospho-Ser at the -1 position in MCC is not involved in major contacts with Scribble PDZ1, and in conjunction with our affinity measurements suggest that the impact of phosphorylation at the -1 position of MCC must extend beyond a simple modulation of the affinity for Scribble PDZ domains.

Does bathymetry drive coastal whale shark (Rhincodon typus) aggregations?

BACKGROUND: The whale shark (Rhincodon typus) is known to aggregate in a number of coastal locations globally, however what causes these aggregations to form where they do is largely unknown. This study examines whether bathymetry is an important driver of coastal aggregation locations for R. typus through bathymetry's effect on primary productivity and prey availability. This is a global study taking into account all coastal areas within R. typus' range. METHODS: R. typus aggregation locations were identified through an extensive literature review. Global bathymetric data were compared at R. typus aggregation locations and a large random selection of non-aggregation areas. Generalised linear models were used to assess which bathymetric characteristic had the biggest influence on aggregation presence. RESULTS: Aggregation sites were significantly shallower than non-aggregation sites and in closer proximity to deep water (the mesopelagic zone) by two orders of magnitude. Slope at aggregation sites was significantly steeper than non-aggregation sites. These three bathymetric variables were shown to have the biggest association with aggregation sites, with up to 88% of deviation explained by the GLMs. DISCUSSION: The three key bathymetric characteristics similar at the aggregation sites are known to induce upwelling events, increase primary productivity and consequently attract numerous other filter feeding species. The location of aggregation sites in these key areas can be attributed to this increased prey availability, thought to be the main reason R. typus aggregations occur, extensively outlined in the literature. The proximity of aggregations to shallow areas such as reefs could also be an important factor why whale sharks thermoregulate after deep dives to feed. These findings increase our understanding of whale shark behaviour and may help guide the identification and conservation of further aggregation sites.

Drosophila melanogaster Guk-holder interacts with the Scribbled PDZ1 domain and regulates epithelial development with Scribbled and Discs Large.

Epithelial cell polarity is controlled by components of the Scribble polarity module, and its regulation is critical for tissue architecture and cell proliferation and migration. In Drosophila melanogaster, the adaptor protein Guk-holder (Gukh) binds to the Scribbled (Scrib) and Discs Large (Dlg) components of the Scribble polarity module and plays an important role in the formation of neuromuscular junctions. However, Gukh's role in epithelial tissue formation and the molecular basis for the Scrib-Gukh interaction remain to be defined. We now show using isothermal titration calorimetry that the Scrib PDZ1 domain is the major site for an interaction with Gukh. Furthermore, we defined the structural basis of this interaction by determining the crystal structure of the Scrib PDZ1-Gukh complex. The C-terminal PDZ-binding motif of Gukh is located in the canonical ligand-binding groove of Scrib PDZ1 and utilizes an unusually extensive network of hydrogen bonds and ionic interactions to enable binding to PDZ1 with high affinity. We next examined the role of Gukh along with those of Scrib and Dlg in Drosophila epithelial tissues and found that Gukh is expressed in larval-wing and eye-epithelial tissues and co-localizes with Scrib and Dlg at the apical cell cortex. Importantly, we show that Gukh functions with Scrib and Dlg in the development of Drosophila epithelial tissues, with depletion of Gukh enhancing the eye- and wing-tissue defects caused by Scrib or Dlg depletion. Overall, our findings reveal that Scrib's PDZ1 domain functions in the interaction with Gukh and that the Scrib-Gukh interaction has a key role in epithelial tissue development in Drosophila.

Sessile and mobile components of a benthic ecosystem display mixed trends within a temperate marine reserve.

Despite recent efforts to increase the global coverage of marine protected areas (MPAs), studies investigating the effectiveness of marine protected areas within temperate waters remain scarce. Furthermore, out of the few studies published on MPAs in temperate waters, the majority focus on specific ecological or fishery components rather than investigating the ecosystem as a whole. This study therefore investigated the dynamics of both benthic communities and fish populations within a recently established, fully protected marine reserve in Lamlash Bay, Isle of Arran, United Kingdom, over a four year period. A combination of photo and diver surveys revealed live maerl (Phymatolithon calcareum), macroalgae, sponges, hydroids, feather stars and eyelash worms (Myxicola infundibulum) to be significantly more abundant within the marine reserve than on surrounding fishing grounds. Likewise, the overall composition of epifaunal communities in and outside the reserve was significantly different. Both results are consistent with the hypothesis that protecting areas from fishing can encourage seafloor habitats to recover. In addition, the greater abundance of complex habitats within the reserve appeared to providing nursery habitat for juvenile cod (Gadus morhua) and scallops (Pecten maximus and Aequipecten opercularis). In contrast, there was little difference in the abundance of mobile benthic fauna, such as crabs and starfish, between the reserve and outside. Similarly, the use of baited underwater video cameras revealed no difference in the abundance and size of fish between the reserve and outside. Limited recovery of these ecosystem components may be due to the relatively small size (2.67 km(2)) and young age of the reserve (<5 years), both of which might have limited the extent of any benefits afforded to mobile fauna and fish communities. Overall, this study provides evidence that fully protected marine reserves can encourage seafloor habitats to recover, which in turn, can create a number of benefits that flow back to other species, including those of commercial importance.