Search for: All records

ABSTRACT Systemic viral infection of insects typically begins with the primary infection of midgut epithelial cells (enterocytes) and subsequent transit of the progeny virus in an apical-to-basal orientation into the hemocoel. For insect-vectored viruses, an oppositely oriented process (basal-to-apical transit) occurs upon secondary infection of salivary glands and is necessary for virus transmission to non-insect hosts. To examine this inversely oriented virus transit in these polarized tissues, we assessed the intracellular trafficking of two model viral envelope proteins (baculovirus GP64 and vesicular stomatitis virus G) in the midgut and salivary gland cells of the model insect,Drosophila melanogaster. Using fly lines that inducibly express either GP64 or VSV G, we found that each protein, expressed alone, was trafficked basally in midgut enterocytes. In salivary gland cells, VSV G was trafficked apically in most but not all cells, whereas GP64 was consistently trafficked basally. We demonstrated that a YxxØ motif present in both proteins was critical for basal trafficking in midgut enterocytes but dispensable for trafficking in salivary gland cells. Using RNAi, we found that clathrin adaptor protein complexes AP-1 and AP-3, as well as seven Rab GTPases, were involved in polarized VSV G trafficking in midgut enterocytes. Our results indicate that these viral envelope proteins encode the requisite information and require no other viral factors for appropriately polarized trafficking. In addition, they exploit tissue-specific differences in protein trafficking pathways to facilitate virus egress in the appropriate orientation for establishing systemic infections and vectoring infection to other hosts. IMPORTANCEViruses that use insects as hosts must navigate specific routes through different insect tissues to complete their life cycles. The routes may differ substantially depending on the life cycle of the virus. Both insect pathogenic viruses and insect-vectored viruses must navigate through the polarized cells of the midgut epithelium to establish a systemic infection. In addition, insect-vectored viruses must also navigate through the polarized salivary gland epithelium for transmission. Thus, insect-vectored viruses appear to traffic in opposite directions in these two tissues. In this study, we asked whether two viral envelope proteins (VSV G and baculovirus GP64) alone encode the signals necessary for the polarized trafficking associated with their respective life cycles. UsingDrosophilaas a model to examine tissue-specific polarized trafficking of these viral envelope proteins, we identified one of the virus-encoded signals and several host proteins associated with regulating the polarized trafficking in the midgut epithelium. 

ABSTRACT The baculovirus envelope protein GP64 is an essential component of the budded virus and is necessary for efficient virion assembly. Little is known regarding intracellular trafficking of GP64 to the plasma membrane, where it is incorporated into budding virions during egress. To identify host proteins and potential cellular trafficking pathways that are involved in delivery of GP64 to the plasma membrane, we developed and characterized a stable Drosophila cell line that inducibly expresses the AcMNPV GP64 protein and used that cell line in combination with a targeted RNA interference (RNAi) screen of vesicular protein trafficking pathway genes. Of the 37 initial hits from the screen, we validated and examined six host genes that were important for trafficking of GP64 to the cell surface. Validated hits included Rab GTPases Rab1 and Rab4 , Clathrin heavy chain , clathrin adaptor protein genes AP-1-2β and AP-2μ , and Snap29 . Two gene knockdowns ( Rab5 and Exo84 ) caused substantial increases (up to 2.5-fold) of GP64 on the plasma membrane. We found that a small amount of GP64 is released from cells in exosomes and that some portion of cell surface GP64 is endocytosed, suggesting that recycling helps to maintain GP64 at the cell surface. IMPORTANCE While much is known regarding trafficking of viral envelope proteins in mammalian cells, little is known about this process in insect cells. To begin to understand which factors and pathways are needed for trafficking of insect virus envelope proteins, we engineered a Drosophila melanogaster cell line and implemented an RNAi screen to identify cellular proteins that aid transport of the model baculovirus envelope protein (GP64) to the cell surface. For this we developed an experimental system that leverages the large array of tools available for Drosophila and performed a targeted RNAi screen to identify cellular proteins involved in GP64 trafficking to the cell surface. Since viral envelope proteins are often critical for production of infectious progeny virions, these studies lay the foundation for understanding how either pathogenic insect viruses (baculoviruses) or insect-vectored viruses (e.g., flaviviruses, alphaviruses) egress from cells in tissues such as the midgut to enable systemic virus infection. 

One of the most well-known extragalactic sources in the sky, quasar3C 454.3, shows a curved parsec-scale jet that has been exhaustively monitored with very-long-baseline interferometry (VLBI) over the recent years. In this work, we present a comprehensive analysis of four years of high-frequency VLBI observations at 43 GHz and 86 GHz, between 2013–2017, in total intensity and linear polarization. The images obtained from these observations enabled us to study the jet structure and the magnetic field topology of the source on spatial scales down to 4.6 parsec in projected distance. The kinematic analysis reveals the abrupt vanishing of at least four new superluminal jet features in a characteristic jet region (i.e., region C), which is located at an approximate distance of 0.6 milliarcsec from the VLBI core. Our results support a model in which the jet bends, directing the relativistic plasma flow almost perfectly toward our line of sight, co-spatially with the region where components appear to stop. 

Context. Because of its proximity and the large size of its black hole, M 87 is one of the best targets for studying the launching mechanism of active galactic nucleus jets. Currently, magnetic fields are considered to be an essential factor in the launching and accelerating of the jet. However, current observational estimates of the magnetic field strength of the M 87 jet are limited to the innermost part of the jet (≲100 r s ) or to HST-1 (∼10 5   r s ). No attempt has yet been made to measure the magnetic field strength in between. Aims. We aim to infer the magnetic field strength of the M 87 jet out to a distance of several thousand r s by tracking the distance-dependent changes in the synchrotron spectrum of the jet from high-resolution very long baseline interferometry observations. Methods. In order to obtain high-quality spectral index maps, quasi-simultaneous observations at 22 and 43 GHz were conducted using the KVN and VERA Array (KaVA) and the Very Long Baseline Array (VLBA). We compared the spectral index distributions obtained from the observations with a model and placed limits on the magnetic field strengths as a function of distance. Results. The overall spectral morphology is broadly consistent over the course of these observations. The observed synchrotron spectrum rapidly steepens from α 22 − 43 GHz  ∼ −0.7 at ∼2 mas to α 22 − 43 GHz  ∼ −2.5 at ∼6 mas. In the KaVA observations, the spectral index remains unchanged until ∼10 mas, but this trend is unclear in the VLBA observations. A spectral index model in which nonthermal electron injections inside the jet decrease with distance can adequately reproduce the observed trend. This suggests the magnetic field strength of the jet at a distance of 2−10 mas (∼900 r s  − ∼4500 r s in the deprojected distance) has a range of B  = (0.3−1.0 G)( z /2mas) −0.73 . Extrapolating to the Event Horizon Telescope scale yields consistent results, suggesting that the majority of the magnetic flux of the jet near the black hole is preserved out to ∼4500 r s without significant dissipation. 

The East Asian VLBI Network (EAVN) is an international VLBI facility in East Asia and is operated under mutual collaboration between East Asian countries, as well as part of Southeast Asian and European countries. EAVN currently consists of 16 radio telescopes and three correlators located in China, Japan, and Korea, and is operated mainly at three frequency bands, 6.7, 22, and 43 GHz with the longest baseline length of 5078 km, resulting in the highest angular resolution of 0.28 milliarcseconds at 43 GHz. One of distinct capabilities of EAVN is multi-frequency simultaneous data reception at nine telescopes, which enable us to employ the frequency phase transfer technique to obtain better sensitivity at higher observing frequencies. EAVN started its open-use program in the second half of 2018, providing a total observing time of more than 1100 h in a year. EAVN fills geographical gap in global VLBI array, resulting in enabling us to conduct contiguous high-resolution VLBI observations. EAVN has produced various scientific accomplishments especially in observations toward active galactic nuclei, evolved stars, and star-forming regions. These activities motivate us to initiate launch of the ’Global VLBI Alliance’ to provide an opportunity of VLBI observation with the longest baselines on the earth. 

Abstract Sagittarius A* (Sgr A*), the Galactic Center supermassive black hole (SMBH), is one of the best targets in which to resolve the innermost region of an SMBH with very long baseline interferometry (VLBI). In this study, we have carried out observations toward Sgr A* at 1.349 cm (22.223 GHz) and 6.950 mm (43.135 GHz) with the East Asian VLBI Network, as a part of the multiwavelength campaign of the Event Horizon Telescope (EHT) in 2017 April. To mitigate scattering effects, the physically motivated scattering kernel model from Psaltis et al. (2018) and the scattering parameters from Johnson et al. (2018) have been applied. As a result, a single, symmetric Gaussian model well describes the intrinsic structure of Sgr A* at both wavelengths. From closure amplitudes, the major-axis sizes are ∼704 ± 102μas (axial ratio ∼ ${1.19}_{-0.19}^{+0.24}$) and ∼300 ± 25μas (axial ratio ∼1.28 ± 0.2) at 1.349 cm and 6.95 mm, respectively. Together with a quasi-simultaneous observation at 3.5 mm (86 GHz) by Issaoun et al. (2019), we show that the intrinsic size scales with observing wavelength as a power law, with an index ∼1.2 ± 0.2. Our results also provide estimates of the size and compact flux density at 1.3 mm, which can be incorporated into the analysis of the EHT observations. In terms of the origin of radio emission, we have compared the intrinsic structures with the accretion flow scenario, especially the radiatively inefficient accretion flow based on the Keplerian shell model. With this, we show that a nonthermal electron population is necessary to reproduce the source sizes.