Analysis of Endocytic Pathways in Drosophila Cells Reveals a Conserved Role for GBF1 in Internalization via GEECs

In mammalian cells, endocytosis of the fluid phase and glycosylphosphatidylinositol-anchored proteins (GPI-APs) forms GEECs (GPI-AP enriched early endosomal compartments) via an Arf1- and Cdc42-mediated, dynamin independent mechanism. Here we use four different fluorescently labeled probes and several markers in combination with quantitative kinetic assays, RNA interference and high resolution imaging to delineate major endocytic routes in Drosophila cultured cells. We find that the hallmarks of the pinocytic GEEC pathway are conserved in Drosophila and identify garz, the fly ortholog of the GTP exchange factor GBF1, as a novel component of this pathway. Live confocal and TIRF imaging reveals that a fraction of GBF1 GFP dynamically associates with ABD RFP (a sensor for activated Arf1 present on nascent pinosomes). Correspondingly, a GTP exchange mutant of GBF1 has altered ABD RFP localization in the evanescent field and is impaired in fluid phase uptake. Furthermore, GBF1 activation is required for the GEEC pathway even in the presence of Brefeldin A, implying that, like Arf1, it has a role in endocytosis that is separable from its role in secretion

Observation of B-s(0) -> (D)over-bar*(0)phi and search for B-0 -> (D)over-bar(0)phi decays

The first observation of the $B_s^0 \to \overline{D}^{*0} \phi$ decay is reported, with a significance of more than seven standard deviations, from an analysis of $pp$ collision data corresponding to an integrated luminosity of 3 fb$^{-1}$, collected with the LHCb detector at centre-of-mass energies of $7$ and $8$ TeV. The branching fraction is measured relative to that of the topologically similar decay $B^0 \to \overline{D}^0 \pi^+\pi^-$ and is found to be $\mathcal{B}(B_s^0 \to \overline{D}^{*0} \phi) = (3.7 \pm 0.5 \pm 0.3 \pm 0.2) \times 10^{-5}$, where the first uncertainty is statistical, the second systematic, and the third from the branching fraction of the $B^0 \to \overline{D}^0 \pi^+\pi^-$ decay. The fraction of longitudinal polarisation in this decay is measured to be ${f_{\rm L} =(73 \pm 15 \pm 3)\%}$. The most precise determination of the branching fraction for the $B_s^0 \to \overline{D}^{0} \phi$ decay is also obtained, $\mathcal{B}(B_s^0 \to \overline{D}^{0} \phi) = (3.0 \pm 0.3 \pm 0.2 \pm 0.2) \times 10^{-5}$. An upper limit, $\mathcal{B}(B^0 \to \overline{D}^{0} \phi) < 2.0 \ (2.2) \times 10^{-6}$ at $90\%$ (95\%) confidence level is set. A constraint on the $\omega-\phi$ mixing angle $\delta$ is set at $|\delta| < 5.2^\circ~ (5.5^\circ)$ at $90\%$ ($95\%$) confidence level.Comment: All figures and tables, along with any supplementary material and additional information, are available at https://lhcbproject.web.cern.ch/lhcbproject/Publications/LHCbProjectPublic/LHCb-PAPER-2018-015.htm