Information seeking behavioural paths of physicians for diabetes mellitus care: a qualitative comparative analysis of information needs, sources, and barriers

This study addresses diabetes physicians’ information seeking behavioural paths (digital, conventional, interpersonal) which lead to information needs satisfaction and the barriers encountered in this process. The study was based on empirical evidence from a survey of 159 physicians. Theoretical analysis was informed by Wilson’s model of information seeking behaviour. The data were analysed using fuzzy set qualitative comparative analysis method. The method was successful in identifying five behavioural paths leading to physicians’ information needs satisfaction (professional/health coaching) which demonstrate different relationships between information sources (conventional/interpersonal/digital) and information barriers (personal/digital illiteracy) and five behavioural paths that are not leading to satisfaction

Transporter membrane traffic and function: lessons from a mould

Transporters are essential transmembrane proteins that mediate the selective translocation of solutes, ions or drugs across biological membranes. Their function is related to cell nutrition, communication, stress resistance and homeostasis. Consequently, their malfunction is associated with genetic or metabolic diseases and drug sensitivity or resistance. A distinctive characteristic of transporters is their cotranslational translocation and folding in a membrane bilayer, this being the endoplasmic reticulum (ER) in eukaryotes or the cell membrane in prokaryotes. In the former case, transporters exit the ER packed in secretory vesicles and traffic via seemingly unconventional, rather than Golgi-dependent, sorting routes to their final destination, the plasma membrane (PM). Proper folding is a prerequisite for ER exit and further trafficking. Misfolded transporters, either due to mutations, high temperature of chemical agents (e.g. DMSO, DTT) are blocked in the ER. The accumulation of ER-retained transporters, in most cases, elicits endoplasmic reticulum-associated degradation, but also ubiquitination-dependent, chaperone-mediated, selective autophagy. The function of PM transporters is finely regulated at the cellular level, in response to physiological or stress signals that promote, via α-arrestin-assisted ubiquitination, their endocytosis and vacuolar/lysosomal degradation, and in some cases recycling to the PM. Importantly, transporter oligomerization and specific interactions with membrane lipids are emerging as important players in transporter expression, function and turnover. This review discusses how paradigmatic work on transporters of a model mould, Aspergillus nidulans, has contributed to novel findings related to transporter functioning in eukaryotes. © 2019 Federation of European Biochemical Societie

Secretory vesicle polar sorting, endosome recycling and cytoskeleton organization require the AP-1 complex in aspergillus nidulans

The AP-1 complex is essential for membrane protein traffic via its role in the pinching-off and sorting of secretory vesicles (SVs) from the trans-Golgi and/or endosomes. While its essentiality is undisputed in metazoa, its role in simpler eukaryotes seems less clear. Here, we dissect the role of AP-1 in the filamentous fungus Aspergillus nidulans and show that it is absolutely essential for growth due to its role in clathrin-dependent maintenance of polar traffic of specific membrane cargoes toward the apex of growing hyphae. We provide evidence that AP-1 is involved in both anterograde sorting of RabERab11-labeled SVs and RabA/BRab5-dependent endosome recycling. Additionally, AP-1 is shown to be critical for microtubule and septin organization, further rationalizing its essentiality in cells that face the challenge of cytoskeleton-dependent polarized cargo traffic. This work also opens a novel issue on how nonpolar cargoes, such as transporters, are sorted to the eukaryotic plasma membrane. © 2018 by the Genetics Society of America

Translocation of nutrient transporters to cell membrane via Golgi bypass in Aspergillus nidulans

Nutrient transporters, being polytopic membrane proteins, are believed, but not formally shown, to traffic from their site of synthesis, the ER, to the plasma membrane through Golgi-dependent vesicular trafficking. Here, we develop a novel genetic system to investigate the trafficking of a neosynthesized model transporter, the well-studied UapA purine transporter of Aspergillus nidulans. We show that sorting of neosynthesized UapA to the plasma membrane (PM) bypasses the Golgi and does not necessitate key Rab GTPases, AP adaptors, microtubules or endosomes. UapA PM localization is found to be dependent on functional COPII vesicles, actin polymerization, clathrin heavy chain and the PM t-SNARE SsoA. Actin polymerization proved to primarily affect COPII vesicle formation, whereas the essential role of ClaH seems indirect and less clear. We provide evidence that other evolutionary and functionally distinct transporters of A. nidulans also follow the herein identified Golgi-independent trafficking route of UapA. Importantly, our findings suggest that specific membrane cargoes drive the formation of distinct COPII subpopulations that bypass the Golgi to be sorted non-polarly to the PM, and thus serving house-keeping cell functions. © 2020 The Author

Oligomerization of the UapA Purine Transporter Is Critical for ER-Exit, Plasma Membrane Localization and Turnover

Central to the process of transmembrane cargo trafficking is the successful folding and exit from the ER (endoplasmic reticulum) through packaging in COPII vesicles. Here, we use the UapA purine transporter of Aspergillus nidulans to investigate the role of cargo oligomerization in membrane trafficking. We show that UapA oligomerizes (at least dimerizes) and that oligomerization persists upon UapA endocytosis and vacuolar sorting. Using a validated bimolecular fluorescence complementation assay, we provide evidence that a UapA oligomerization is associated with ER-exit and turnover, as ER-retained mutants due to either modification of a Tyr-based N-terminal motif or partial misfolding physically associate but do not associate properly. Co-expression of ER-retained mutants with wild-type UapA leads to in trans plasma membrane localization of the former, confirming that oligomerization initiates in the ER. Genetic suppression of an N-terminal mutation in the Tyr motif and mutational analysis suggest that transmembrane α-helix 7 affects the oligomerization interface. Our results reveal that transporter oligomerization is essential for membrane trafficking and turnover and is a common theme in fungi and mammalian cells. © 2015 Elsevier Ltd