239 research outputs found
HECT ubiquitin ligases link viral and cellular PPXY motifs to the vacuolar protein-sorting pathway
Many enveloped viruses exploit the class E vacuolar protein-sorting (VPS) pathway to bud from cells, and use peptide motifs to recruit specific class E VPS factors. Homologous to E6AP COOH terminus (HECT) ubiquitin ligases have been implicated as cofactors for PPXY motif–dependent budding, but precisely which members of this family are responsible, and how they access the VPS pathway is unclear. Here, we show that PPXY-dependent viral budding is unusually sensitive to inhibitory fragments derived from specific HECT ubiquitin ligases, namely WWP1 and WWP2. We also show that WWP1, WWP2, or Itch ubiquitin ligase recruitment promotes PPXY-dependent virion release, and that this function requires that the HECT ubiquitin ligase domain be catalytically active. Finally, we show that several mammalian HECT ubiquitin ligases, including WWP1, WWP2, and Itch are recruited to class E compartments induced by dominant negative forms of the class E VPS ATPase, VPS4. These data indicate that specific HECT ubiquitin ligases can link PPXY motifs to the VPS pathway to induce viral budding
HIV-1 Vpu Promotes Release and Prevents Endocytosis of Nascent Retrovirus Particles from the Plasma Membrane
The human immunodeficiency virus (HIV) type-1 viral protein U (Vpu) protein enhances the release of diverse retroviruses from human, but not monkey, cells and is thought to do so by ablating a dominant restriction to particle release. Here, we determined how Vpu expression affects the subcellular distribution of HIV-1 and murine leukemia virus (MLV) Gag proteins in human cells where Vpu is, or is not, required for efficient particle release. In HeLa cells, where Vpu enhances HIV-1 and MLV release approximately 10-fold, concentrations of HIV-1 Gag and MLV Gag fused to cyan fluorescent protein (CFP) were initially detected at the plasma membrane, but then accumulated over time in early and late endosomes. Endosomal accumulation of Gag-CFP was prevented by Vpu expression and, importantly, inhibition of plasma membrane to early endosome transport by dominant negative mutants of Rab5a, dynamin, and EPS-15. Additionally, accumulation of both HIV and MLV Gag in endosomes required a functional late-budding domain. In human HOS cells, where HIV-1 and MLV release was efficient even in the absence of Vpu, Gag proteins were localized predominantly at the plasma membrane, irrespective of Vpu expression or manipulation of endocytic transport. While these data indicated that Vpu inhibits nascent virion endocytosis, Vpu did not affect transferrin endocytosis. Moreover, inhibition of endocytosis did not restore Vpu-defective HIV-1 release in HeLa cells, but instead resulted in accumulation of mature virions that could be released from the cell surface by protease treatment. Thus, these findings suggest that a specific activity that is present in HeLa cells, but not in HOS cells, and is counteracted by Vpu, traps assembled retrovirus particles at the cell surface. This entrapment leads to subsequent endocytosis by a Rab5a- and clathrin-dependent mechanism and intracellular sequestration of virions in endosomes
A Human IgSF Cell-Surface Interactome Reveals a Complex Network of Protein-Protein Interactions
Cell-surface protein-protein interactions (PPIs) mediate cell-cell communication, recognition, and responses. We executed an interactome screen of 564 human cell-surface and secreted proteins, most of which are immunoglobulin superfamily (IgSF) proteins, using a high-throughput, automated ELISA-based screening platform employing a pooled-protein strategy to test all 318,096 PPI combinations. Screen results, augmented by phylogenetic homology analysis, revealed ∼380 previously unreported PPIs. We validated a subset using surface plasmon resonance and cell binding assays. Observed PPIs reveal a large and complex network of interactions both within and across biological systems. We identified new PPIs for receptors with well-characterized ligands and binding partners for “orphan” receptors. New PPIs include proteins expressed on multiple cell types and involved in diverse processes including immune and nervous system development and function, differentiation/proliferation, metabolism, vascularization, and reproduction. These PPIs provide a resource for further biological investigation into their functional relevance and may offer new therapeutic drug targets
A Human IgSF Cell-Surface Interactome Reveals a Complex Network of Protein-Protein Interactions
Cell-surface protein-protein interactions (PPIs) mediate cell-cell communication, recognition, and responses. We executed an interactome screen of 564 human cell-surface and secreted proteins, most of which are immunoglobulin superfamily (IgSF) proteins, using a high-throughput, automated ELISA-based screening platform employing a pooled-protein strategy to test all 318,096 PPI combinations. Screen results, augmented by phylogenetic homology analysis, revealed ∼380 previously unreported PPIs. We validated a subset using surface plasmon resonance and cell binding assays. Observed PPIs reveal a large and complex network of interactions both within and across biological systems. We identified new PPIs for receptors with well-characterized ligands and binding partners for “orphan” receptors. New PPIs include proteins expressed on multiple cell types and involved in diverse processes including immune and nervous system development and function, differentiation/proliferation, metabolism, vascularization, and reproduction. These PPIs provide a resource for further biological investigation into their functional relevance and may offer new therapeutic drug targets
KELT-3b: A Hot Jupiter Transiting a V=9.8 Late-F Star
We report the discovery of KELT-3b, a moderately inflated transiting hot
Jupiter with a mass of 1.477 (-0.067, +0.066) M_J, and radius of 1.345 +/-
0.072 R_J, with an orbital period of 2.7033904 +/- 0.000010 days. The host
star, KELT-3, is a V=9.8 late F star with M_* = 1.278 (-0.061, +0.063) M_sun,
R_* = 1.472 (-0.067, +0.065) R_sun, T_eff = 6306 (-49, +50) K, log(g) = 4.209
(-0.031, +0.033), and [Fe/H] = 0.044 (-0.082, +0.080), and has a likely proper
motion companion. KELT-3b is the third transiting exoplanet discovered by the
KELT survey, and is orbiting one of the 20 brightest known transiting planet
host stars, making it a promising candidate for detailed characterization
studies. Although we infer that KELT-3 is significantly evolved, a preliminary
analysis of the stellar and orbital evolution of the system suggests that the
planet has likely always received a level of incident flux above the
empirically-identified threshold for radius inflation suggested by Demory &
Seager (2011).Comment: 12 pages, 12 figures, accepted to Ap
KELT-8b: A highly inflated transiting hot Jupiter and a new technique for extracting high-precision radial velocities from noisy spectra
We announce the discovery of a highly inflated transiting hot Jupiter
discovered by the KELT-North survey. A global analysis including constraints
from isochrones indicates that the V = 10.8 host star (HD 343246) is a mildly
evolved, G dwarf with K, , , an inferred mass
M, and radius
R. The planetary companion has mass , radius
, surface gravity , and density
g cm. The planet is on a roughly
circular orbit with semimajor axis AU and
eccentricity . The best-fit linear ephemeris is
BJD and
days. This planet is one of the most inflated of all known transiting
exoplanets, making it one of the few members of a class of extremely low
density, highly-irradiated gas giants. The low stellar and large
implied radius are supported by stellar density constraints from follow-up
light curves, plus an evolutionary and space motion analysis. We also develop a
new technique to extract high precision radial velocities from noisy spectra
that reduces the observing time needed to confirm transiting planet candidates.
This planet boasts deep transits of a bright star, a large inferred atmospheric
scale height, and a high equilibrium temperature of
K, assuming zero albedo and perfect heat redistribution, making it one of the
best targets for future atmospheric characterization studies.Comment: Submitted to ApJ, feedback is welcom
KELT-6b: A P~7.9 d Hot Saturn Transiting a Metal-Poor Star with a Long-Period Companion
We report the discovery of KELT-6b, a mildly-inflated Saturn-mass planet
transiting a metal-poor host. The initial transit signal was identified in
KELT-North survey data, and the planetary nature of the occulter was
established using a combination of follow-up photometry, high-resolution
imaging, high-resolution spectroscopy, and precise radial velocity
measurements. The fiducial model from a global analysis including constraints
from isochrones indicates that the V=10.38 host star (BD+31 2447) is a mildly
evolved, late-F star with T_eff=6102 \pm 43 K, log(g_*)=4.07_{-0.07}^{+0.04}
and [Fe/H]=-0.28 \pm 0.04, with an inferred mass M_*=1.09 \pm 0.04 M_sun and
radius R_star=1.58_{-0.09}^{+0.16} R_sun. The planetary companion has mass
M_P=0.43 \pm 0.05 M_J, radius R_P=1.19_{-0.08}^{+0.13} R_J, surface gravity
log(g_P)=2.86_{-0.08}^{+0.06}, and density rho_P=0.31_{-0.08}^{+0.07}
g~cm^{-3}. The planet is on an orbit with semimajor axis a=0.079 \pm 0.001 AU
and eccentricity e=0.22_{-0.10}^{+0.12}, which is roughly consistent with
circular, and has ephemeris of T_c(BJD_TDB)=2456347.79679 \pm 0.00036 and
P=7.845631 \pm 0.000046 d. Equally plausible fits that employ empirical
constraints on the host star parameters rather than isochrones yield a larger
planet mass and radius by ~4-7%. KELT-6b has surface gravity and incident flux
similar to HD209458b, but orbits a host that is more metal poor than HD209458
by ~0.3 dex. Thus, the KELT-6 system offers an opportunity to perform a
comparative measurement of two similar planets in similar environments around
stars of very different metallicities. The precise radial velocity data also
reveal an acceleration indicative of a longer-period third body in the system,
although the companion is not detected in Keck adaptive optics images.Comment: Published in AJ, 17 pages, 15 figures, 6 table
Very Low Mass Stellar and Substellar Companions to Solar-Like Stars From MARVELS V: A Low Eccentricity Brown Dwarf from the Driest Part of the Desert, MARVELS-6b
We describe the discovery of a likely brown dwarf (BD) companion with a
minimum mass of 31.7 +/- 2.0 M_Jup to GSC 03546-01452 from the MARVELS radial
velocity survey, which we designate as MARVELS-6b. For reasonable priors, our
analysis gives a probability of 72% that MARVELS-6b has a mass below the
hydrogen-burning limit of 0.072 M_Sun, and thus it is a high-confidence BD
companion. It has a moderately long orbital period of 47.8929 +0.0063/-0.0062
days with a low eccentricty of 0.1442 +0.0078/-0.0073, and a semi-amplitude of
1644 +12/-13 m/s. Moderate resolution spectroscopy of the host star has
determined the following parameters: T_eff = 5598 +/- 63, log g = 4.44 +/-
0.17, and [Fe/H] = +0.40 +/- 0.09. Based upon these measurements, GSC
03546-01452 has a probable mass and radius of M_star = 1.11 +/- 0.11 M_Sun and
R_star = 1.06 +/- 0.23 R_Sun with an age consistent with less than ~6 Gyr at a
distance of 219 +/- 21 pc from the Sun. Although MARVELS-6b is not observed to
transit, we cannot definitively rule out a transiting configuration based on
our observations. There is a visual companion detected with Lucky Imaging at
7.7 arcsec from the host star, but our analysis shows that it is not bound to
this system. The minimum mass of MARVELS-6b exists at the minimum of the mass
functions for both stars and planets, making this a rare object even compared
to other BDs.Comment: 15 pages, 15 figures, 5 tables. Accepted for publication in The
Astronomical Journa
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