Rab4 Orchestrates a Small GTPase Cascade for Recruitment of Adaptor Proteins to Early Endosomes

SummaryBackgroundEarly, sorting endosomes are a major crossroad of membrane traffic, at the intersection of the endocytic and exocytic pathways. The sorting of endosomal cargo for delivery to different subcellular destinations is mediated by a number of distinct coat protein complexes, including adaptor protein 1 (AP-1), AP-3, and Golgi-localized, gamma adaptin ear-containing, Arf-binding (GGAs) protein. Ultrastructural studies suggest that these coats assemble onto tubular subdomains of the endosomal membrane, but the mechanisms of coat recruitment and assembly at this site remain poorly understood.ResultsHere we report that the endosomal Rab protein Rab4 orchestrates a GTPase cascade that results in the sequential recruitment of the ADP-ribosylation factor (Arf)-like protein Arl1; the Arf-specific guanine nucleotide exchange factors BIG1 and BIG2; and the class I Arfs, Arf1 and Arf3. Knockdown of Arf1, or inhibition of BIG1 and BIG2 activity with brefeldin A results in the loss of AP-1, AP-3, and GGA-3, but not Arl1, from endosomal membranes and the formation of elongated tubules. In contrast, depletion of Arl1 randomizes the distribution of Rab4 on endosomal membranes, inhibits the formation of tubular subdomains, and blocks recruitment of BIG1 and BIG2, Arfs, and adaptor protein complexes to the endosome.ConclusionsTogether these findings indicate that Arl1 links Rab4-dependent formation of endosomal sorting domains with downstream assembly of adaptor protein complexes that constitute the endosomal sorting machinery

High-energy sources at low radio frequency : the Murchison Widefield Array view of Fermi blazars

This is the accepted version of the following article: Giroletti, M. et al., A&A, 588 (2016) A141, which has been published in final form at DOI: http://dx.doi.org/10.1051/0004-6361/201527817. This article may be used for non-commercial purposes in accordance with the EDP Sciences self-archiving policies.Low-frequency radio arrays are opening a new window for the study of the sky, both to study new phenomena and to better characterize known source classes. Being flat-spectrum sources, blazars are so far poorly studied at low radio frequencies. We characterize the spectral properties of the blazar population at low radio frequency compare the radio and high-energy properties of the gamma-ray blazar population, and search for radio counterparts of unidentified gamma-ray sources. We cross-correlated the 6,100 deg^2 Murchison Widefield Array Commissioning Survey catalogue with the Roma blazar catalogue, the third catalogue of active galactic nuclei detected by Fermi-LAT, and the unidentified members of the entire third catalogue of gamma-ray sources detected by \fermilat. When available, we also added high-frequency radio data from the Australia Telescope 20 GHz catalogue. We find low-frequency counterparts for 186 out of 517 (36%) blazars, 79 out of 174 (45%) gamma-ray blazars, and 8 out of 73 (11%) gamma-ray blazar candidates. The mean low-frequency (120--180 MHz) blazar spectral index is $\langle \alpha_\mathrm{low} \rangle=0.57\pm0.02$: blazar spectra are flatter than the rest of the population of low-frequency sources, but are steeper than at $\sim$GHz frequencies. Low-frequency radio flux density and gamma-ray energy flux display a mildly significant and broadly scattered correlation. Ten unidentified gamma-ray sources have a (probably fortuitous) positional match with low radio frequency sources. Low-frequency radio astronomy provides important information about sources with a flat radio spectrum and high energy. However, the relatively low sensitivity of the present surveys still misses a significant fraction of these objects. Upcoming deeper surveys, such as the GaLactic and Extragalactic All-Sky MWA (GLEAM) survey, will provide further insight into this population.Peer reviewedFinal Published versio

Molecular excitation in the Interstellar Medium: recent advances in collisional, radiative and chemical processes

We review the different excitation processes in the interstellar mediumComment: Accepted in Chem. Re

Long-Baseline Neutrino Facility (LBNF) and Deep Underground Neutrino Experiment (DUNE) Conceptual Design Report Volume 2: The Physics Program for DUNE at LBNF

The Physics Program for the Deep Underground Neutrino Experiment (DUNE) at the Fermilab Long-Baseline Neutrino Facility (LBNF) is described

CENP-C recruits M18BP1 to centromeres to promote CENP-A chromatin assembly

CENP-C provides a link between existing CENP-A chromatin and the proteins required for new CENP-A nucleosome assembly

Production and characterization of a recombinant single-chain antibody against Hantaan virus envelop glycoprotein

Hantaan virus (HTNV) is the type of Hantavirus causing hemorrhagic fever with renal syndrome, for which no specific therapeutics are available so far. Cell type-specific internalizing antibodies can be used to deliver therapeutics intracellularly to target cell and thus, have potential application in anti-HTNV infection. To achieve intracellular delivery of therapeutics, it is necessary to obtain antibodies that demonstrate sufficient cell type-specific binding, internalizing, and desired cellular trafficking. Here, we describe the prokaryotic expression, affinity purification, and functional testing of a single-chain Fv antibody fragment (scFv) against HTNV envelop glycoprotein (GP), an HTNV-specific antigen normally located on the membranes of HTNV-infected cells. This HTNV GP-targeting antibody, scFv3G1, was produced in the cytoplasm of Escherichia coli cells as a soluble protein and was purified by immobilized metal affinity chromatography. The purified scFv possessed a high specific antigen-binding activity to HTNV GP and HTNV-infected Vero E6 cells and could be internalized into HTNV-infected cells probably through the clathrin-dependent endocytosis pathways similar to that observed with transferrin. Our results showed that the E. coli-produced scFv had potential applications in targeted and intracellular delivery of therapeutics against HTNV infections

Stereodivergent Synthesis of Enantioenriched 4-Hydroxy-2- cyclopentenones

Protected 4-hydroxycyclopentenones (4-HCPs) constitute an important class of intermediates in chemical synthesis. A route to this class of compound has been developed. Key steps include Noyori reduction (which establishes the stereochemistry of the product), ring-closing metathesis, and simple functional group conversions to provide a set of substituted 4-HCPs in either enantiomeric form

Power spectrum analysis of ionospheric fluctuations with the Murchison Widefield Array

Low-frequency, wide field-of-view (FOV) radio telescopes such as the Murchison Widefield Array (MWA) enable the ionosphere to be sampled at high spatial completeness. We present the results of the first power spectrum analysis of ionospheric fluctuations in MWA data, where we examined the position offsets of radio sources appearing in two data sets. The refractive shifts in the positions of celestial sources are proportional to spatial gradients in the electron column density transverse to the line of sight. These can be used to probe plasma structures and waves in the ionosphere. The regional (10–100 km) scales probed by the MWA, determined by the size of its FOV and the spatial density of radio sources (typically thousands in a single FOV), complement the global (100–1000 km) scales of GPS studies and local (0.01–1 km) scales of radar scattering measurements. Our data exhibit a range of complex structures and waves. Some fluctuations have the characteristics of traveling ionospheric disturbances, while others take the form of narrow, slowly drifting bands aligned along the Earth's magnetic field

New Frontiers-class Uranus Orbiter: Exploring the feasibility of achieving multidisciplinary science with a mid-scale mission

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