Membrane Tethering Complexes in the Endosomal System

Vesicles that are generated by endocytic events at the plasma membrane are destined to early endosomes. A prerequisite for proper fusion is the tethering of two membrane entities. Tethering of vesicles to early endosomes is mediated by the CORVET complex, while fusion of late endosomes with lysosomes depends on the HOPS complex. Recycling through the TGN and to the plasma membrane is facilitated by the GARP and EARP complexes, respectively. However, there are other tethering functions in the endosomal system as there are multiple pathways through which proteins can be delivered from endosomes to either the TGN or the plasma membrane. Furthermore, complexes that may be part of novel tethering complexes have been recently identified. Thus it is likely that more tethering factors exist. In this review, I will provide an overview of different tethering complexes of the endosomal system and discuss how they may provide specificity in membrane traffic

On the fate of early endosomes

Proteins are endocytosed by various pathways into the cell. All these pathways converge at the level of the early endosome. The fate of the early endosome and how proteins are sorted into recycling and late endosomes/multi-vesicular body is a matter of debate and intense research. Obviously, the transition from early to late endosome poses an interesting logistic problem and would merit attention on an intellectual level. Numerous diseases are also caused by defects in turning off/over signaling molecules or mis-sorting of proteins at the level of the early endosome. This brief review aims to discuss different molecular mechanisms whereby early-to-late endosome transition could be achieve

The endoplasmic reticulum-the caring mother of the cell

In eukaryotic cells, various cellular functions are compartmentalized and performed by sophisticated and specialized organelles. However, the membrane-bounded organelles need to communicate with each other and with the cytoplasm, and sense the outside through the plasma membrane to coordinate various functions and to maintain cellular homeostasis. To maintain homeostasis, the information on the cellular state must be collected and appropriate responses initiated. The endoplasmic reticulum fulfils these functions. In this review, I will discuss various aspects of how the ER senses and relays information and acts to protect the cell, in what sometimes could be interpreted as an altruistic behavior

Über die Rolle der Endozytose bei Krankheiten

Zusammenfassung: Endozytose beschreibt einen vielschichtigen, evolutionär konservierten Prozess, über den eine Vielzahl von Stoffen in eine Zelle aufgenommen und anschließend durch Sortieren an ihren Zielort gelang

Attenuation of N-glycosylation causes polarity and adhesion defects in the C. elegans embryo

The C. elegans early embryo is highly polarized, requiring sequestration of cytoplasmic polarity factors at the plasma membrane. This compartmentalization aids asymmetric distribution of lipids and proteins, which is partially responsible for the fates of the daughter cells. Since most plasma membrane proteins are glycosylated, we determined the effect of N-glycosylation attenuation on cell polarity. While polarity establishment was not perturbed, the AB/P1 size ratio was more variable in embryos with reduced N-glycosylation than in the mock-treated ones. In addition, among other deficiencies, we observed spindle orientation defects in two-cell embryos. Moreover, cell-cell adhesion was specifically lost at the two-cell stage when N-glycosylation was reduced. This loss-of-adhesion phenotype was rescued by interfering with polarity establishment, indicating that polarity establishment enforces plasma membrane compartmentalization. Consistently, the decreased plasma membrane levels of the adhesion proteins E-cadherin and MAGI-1 in ribo-1(RNAi) embryos were restored in the absence of functional PAR-2. Our data suggest a general role for N-glycosylation in plasma membrane compartmentalization and cell polarity

Decapping complex is essential for functional P-body formation and is buffered by nuclear localization

mRNA decay is a key step in regulating the cellular proteome. Cytoplasmic mRNA is largely turned over in processing bodies (P-bodies). P-body units assemble to form P-body granules under stress conditions. How this assembly is regulated, however, remains still poorly understood. Here, we show that the translational repressor Scd6 and the decapping stimulator Edc3 act partially redundantly in P-body assembly by capturing the Dcp1/2 decapping complex and preventing it from becoming imported into the nucleus by the karyopherin ß Kap95. Nuclear Dcp1/2 does not drive mRNA decay and might be stored there as a ready releasable pool, indicating a dynamic equilibrium between cytoplasmic and nuclear Dcp1/2. Cytoplasmic Dcp1/2 is linked to Dhh1 via Edc3 and Scd6. Functional P-bodies are present at the endoplasmic reticulum where Dcp2 potentially acts to increase the local concentration of Dhh1 through interaction with Scd6 and Edc3 to drive phase separation and hence P-body formation

Reconstitution of Retrograde Transport from the Golgi to the ER In Vitro

Retrograde transport from the Golgi to the ER is an essential process. Resident ER proteins that escape the ER and proteins that cycle between the Golgi and the ER must be retrieved. The interdependence of anterograde and retrograde vesicle trafficking makes the dissection of both processes difficult in vivo. We have developed an in vitro system that measures the retrieval of a soluble reporter protein, the precursor of the yeast pheromone α-factor fused to a retrieval signal (HDEL) at its COOH terminus (Dean, N., and H.R.B Pelham. 1990. J. Cell Biol. 111:369–377). Retrieval depends on the HDEL sequence; the α-factor precursor, naturally lacking this sequence, is not retrieved. A full cycle of anterograde and retrograde transport requires a simple set of purified cytosolic proteins, including Sec18p, the Lma1p complex, Uso1p, coatomer, and Arf1p. Among the membrane-bound v-SNAP receptor (v-SNARE) proteins, Bos1p is required only for forward transport, Sec22p only for retrograde trafficking, and Bet1p is implicated in both avenues of transport. Putative retrograde carriers (COPI vesicles) generated from Golgi-enriched membranes contain v-SNAREs as well as Emp47p as cargo

Protein sorting from endosomes to the TGN

Retrograde transport from endosomes to the trans-Golgi network is essential for recycling of protein and lipid cargoes to counterbalance anterograde membrane traffic. Protein cargo subjected to retrograde traffic include lysosomal acid-hydrolase receptors, SNARE proteins, processing enzymes, nutrient transporters, a variety of other transmembrane proteins, and some extracellular non-host proteins such as viral, plant, and bacterial toxins. Efficient delivery of these protein cargo molecules depends on sorting machineries selectively recognizing and concentrating them for their directed retrograde transport from endosomal compartments. In this review, we outline the different retrograde transport pathways governed by various sorting machineries involved in endosome-to-TGN transport. In addition, we discuss how this transport route can be analyzed experimentally

Meeting report - An online gathering about the latest on Molecular Membrane Biology

Gordon Research Conferences (GRC) are among the meetings that are particularly important as they are rather small and foster scientific exchange. Many established researchers participate and meet for a week, often in some remote location. The 2021 molecular membrane biology Gordon conference, usually held at Proctor Academy (New Hampshire, USA) was one of the many meetings affected when the SARS-CoV2 pandemic resulted in an immediate shut down of all public life and world travel. Scientists and scientific organizations reacted immediately and discovered the advantages of online meetings. Several members of the membrane biology community felt that a complete halt of scientific conferences would particularly harm young investigators in their career path as they would not get the chance to present their data in front of the scientific community. As a result, we decided to take the initiative and try to find a format for an online meeting in 2021. Before starting, we sent around a survey to ask for interest, and received overwhelming enthusiasm and support – a testament to the vitality of the membrane trafficking community. We agreed on a format that would mimic somehow the in-person meeting: poster sessions, followed by a dense set of talks with sufficient time for discussion, a meet-the-speaker slot, then another set of talks and eventually another poster session or meeting time. Talks were presented mostly by early career researchers, postdocs or assistant professors. To foster discussion and presentation of unpublished data, talks were live and not recorded. Gather.town was used as a platform for poster sessions and social interactions. Very importantly, the meeting was completely free as funding was secured via four journals (Journal of Cell Biology, Journal of Cell Science, FEBS Journal and Traffic Journal) and three German DFG-funded research consortia (SFB 944, 958 and TR 186), which covered the user fees for all participants