Nbr1 Is a Novel Inhibitor of Ligand-Mediated Receptor Tyrosine Kinase Degradation

endocytic trafficking and selective autophagy. However, the exact function of Nbr1 in these contexts has not been studied in detail. Here we investigated the role of Nbr1 in the trafficking of receptor tyrosine kinases (RTKs). We report that ectopic Nbr1 expression inhibits the ligand-mediated lysosomal degradation of RTKs, and this is probably done via the inhibition of receptor internalization. Conversely, the depletion of endogenous NBR1 enhances RTK degradation. Analyses of truncation mutations demonstrated that the C terminus of Nbr1 is essential but not sufficient for this activity. Moreover, the C terminus of Nbr1 is essential but not sufficient for the localization of the protein to late endosomes. We demonstrate that the C terminus of Nbr1 contains a novel membrane-interacting amphipathic -helix, which is essential for the late endocytic localization of the protein but not for its effect on RTK degradation. Finally, autophagic and late endocytic localizations of Nbr1 are independent of one another, suggesting that the roles of Nbr1 in each context might be distinct. Our results define Nbr1 as a negative regulator of ligand-mediated RTK degradation and reveal the interplay between its various regions for protein localization and function

DNA compaction by the higher-order assembly of PRH/Hex homeodomain protein oligomers

Protein self-organization is essential for the establishment and maintenance of nuclear architecture and for the regulation of gene expression. We have shown previously that the Proline-Rich Homeodomain protein (PRH/Hex) self-assembles to form oligomeric complexes that bind to arrays of PRH binding sites with high affinity and specificity. We have also shown that many PRH target genes contain suitably spaced arrays of PRH sites that allow this protein to bind and regulate transcription. Here, we use analytical ultracentrifugation and electron microscopy to further characterize PRH oligomers. We use the same techniques to show that PRH oligomers bound to long DNA fragments self-associate to form highly ordered assemblies. Electron microscopy and linear dichroism reveal that PRH oligomers can form protein–DNA fibres and that PRH is able to compact DNA in the absence of other proteins. Finally, we show that DNA compaction is not sufficient for the repression of PRH target genes in cells. We conclude that DNA compaction is a consequence of the binding of large PRH oligomers to arrays of binding sites and that PRH is functionally and structurally related to the Lrp/AsnC family of proteins from bacteria and archaea, a group of proteins formerly thought to be without eukaryotic equivalents

DNA compaction by the higher-order assembly of PRH/Hex homeodomain protein oligomers

Protein self-organization is essential for the establishment and maintenance of nuclear architecture and for the regulation of gene expression. We have shown previously that the Proline-Rich Homeodomain protein (PRH/Hex) self-assembles to form oligomeric complexes that bind to arrays of PRH binding sites with high affinity and specificity. We have also shown that many PRH target genes contain suitably spaced arrays of PRH sites that allow this protein to bind and regulate transcription. Here, we use analytical ultracentrifugation and electron microscopy to further characterize PRH oligomers. We use the same techniques to show that PRH oligomers bound to long DNA fragments self-associate to form highly ordered assemblies. Electron microscopy and linear dichroism reveal that PRH oligomers can form protein–DNA fibres and that PRH is able to compact DNA in the absence of other proteins. Finally, we show that DNA compaction is not sufficient for the repression of PRH target genes in cells. We conclude that DNA compaction is a consequence of the binding of large PRH oligomers to arrays of binding sites and that PRH is functionally and structurally related to the Lrp/AsnC family of proteins from bacteria and archaea, a group of proteins formerly thought to be without eukaryotic equivalents

Facilitation of non-indigenous ascidian by marine eco-engineering interventions at an urban site

Marine artificial structures often support lower native species diversity and more non-indigenous species (NIS), but adding complex habitat and using bioreceptive materials have the potential to mitigate these impacts. Here, the interacting effects of structural complexity (flat, complex with pits) and concrete mixture (standard, or with oyster shell or vermiculite aggregate) on recruitment were assessed at two intertidal levels at an urban site. Complex tiles had less green algal cover, oyster shell mixtures had less brown (Ralfsia sp.) algal cover. At a low tidal elevation, the non-indigenous ascidian Styela plicata dominated complex tiles. Additionally, mixtures with oyster shell supported higher total cover of sessile species, and a higher cover of S. plicata. There were no effects of complexity or mixture on biofilm communities and native and NIS richness. Overall, these results suggest that habitat complexity and some bioreceptive materials may facilitate colonisation by a dominant invertebrate invader on artificial structures

Analysis of SMALP co-extracted phospholipids shows distinct membrane environments for three classes of bacterial membrane protein

Biological characterisation of membrane proteins lags behind that of soluble proteins. This reflects issues with the traditional use of detergents for extraction, as the surrounding lipids are generally lost, with adverse structural and functional consequences. In contrast, styrene maleic acid (SMA) copolymers offer a detergent-free method for biological membrane solubilisation to produce SMA-lipid particles (SMALPs) containing membrane proteins together with their surrounding lipid environment. We report the development of a reverse-phase LC-MS/MS method for bacterial phospholipids and the first comparison of the profiles of SMALP co-extracted phospholipids from three exemplar bacterial membrane proteins with different topographies: FtsA (associated membrane protein), ZipA (single transmembrane helix), and PgpB (integral membrane protein). The data showed that while SMA treatment per se did not preferentially extract specific phospholipids from the membrane, SMALP-extracted ZipA showed an enrichment in phosphatidylethanolamines and depletion in cardiolipins compared to the bulk membrane lipid. Comparison of the phospholipid profiles of the 3 SMALP-extracted proteins revealed distinct lipid compositions for each protein: ZipA and PgpB were similar, but in FtsA samples longer chain phosphatidylglycerols and phosphatidylethanolamines were more abundant. This method offers novel information on the phospholipid interactions of these membrane proteins

Identifying the physical features of marina infrastructure associated with the presence of non-native species in the UK

Marine invasive non-native species (NNS) are one of the greatest threats to global marine biodiversity, causing significant economic and social impacts. Marinas are increasingly recognised as key reservoirs for invasive NNS. They provide submersed artificial habitat that unintentionally supports the establishment of NNS introduced from visiting recreational vessels. While ballast water and shipping vectors have been well documented, the role of recreational vessels in spreading NNS has been relatively poorly studied. Identification of the main physical features found within marinas, which relate to the presence of NNS, is important to inform the development of effective biosecurity measures and prevent further spread. Towards this aim, physical features that could influence the presence of NNS were assessed for marinas throughout the UK in July 2013. Thirty-three marine and brackish NNS have been recorded in UK marinas, and of the 88 marinas studied in detail, 83 contained between 1 and 13 NNS. Significant differences in freshwater input, marina entrance width and seawall length were associated with the presence of NNS. Additionally, questionnaires were distributed to marina managers and recreational vessel owners to understand current biosecurity practices and attitudes to recreational vessel biosecurity. The main barriers to biosecurity compliance were cited as cost and time. Further work identifying easily distinguished features of marinas could be used as a proxy to assess risk of invasion. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00227-016-2941-8) contains supplementary material, which is available to authorized users

The hunter River estuary water quality model

© Australasian Coasts and Ports 2019 Conference. All rights reserved. This paper presents a detailed hydrodynamic and water quality model to simulate ecological processes in the Hunter River estuary. Following an extensive 3-year multi-disciplinary field campaign, the model was developed to assess total catchment management options. The model outcomes are linked to existing water sharing plans, pollution reduction plans and coastal reforms underway in NSW. Initially a detailed scoping study was undertaken to determine the values and requirements of the key stakeholders across the catchment. Data gaps were subsequently prioritised, and an inter-agency modelling oversight committee was formed to ensure that the modelling tools would be accepted across the region. Following these developmental stages, a field program was initiated which included: estuary wide flow gauging and water quality assessments, microbial linkages, ecotoxicological assessments, sedimentation dynamics, DNA sequencing, qPCR analyses, catchment hydrological flux measurements, nutrient mesocosm experiments, bathymetry surveys and the development of crop irrigation modules. The field data analyses resulted in a conceptual model of the eco-hydraulics of the estuary. A robust numerical model was formulated through an extensive process of external peer review. A source model was selected that ensured the broadest flexibility and ongoing usage rates. A multi-disciplinary approach was undertaken to ensure the model represents a wide range of estuarine processes. The final model is currently undergoing additional peer review, calibration/validation and simulation testing