V accessory proteins from Paramyxoviruses are important in viral evasion of the innate immune response. Here, using a cell survival assay that identifies both inhibitors and activators of interferon regulatory factor 3 (IRF3)-mediated gene induction, we identified select paramyxoviral V proteins that inhibited double-stranded RNA-mediated signaling; these are encoded by mumps virus (MuV), human parainfluenza virus 2 (hPIV2), and parainfluenza virus 5 (PIV5), all members of the genus Rubulavirus. We showed that interaction between V and the IRF3/7 kinases, TRAF family member-associated NFB activator (TANK)-binding kinase 1 (TBK1)/inhibitor of B kinase ⑀ (IKKe), was essential for this inhibition. Indeed, V proteins were phosphorylated directly by TBK1/IKKe, and this, intriguingly, resulted in lowering of the cellular level of V. Thus, it appears that V mimics IRF3 in both its phosphorylation by TBK1/IKKe and its subsequent degradation. Finally, a PIV5 mutant encoding a V protein that could not inhibit IKKe was much more susceptible to the antiviral effects of double-stranded RNA than the wild-type virus. Because many innate immune response signaling pathways, including those initiated by TLR3, TLR4, RIG-I, MDA5, and DNA-dependent activator of IRFs (DAI), use TBK1/ IKKe as the terminal kinases to activate IRFs, rubulaviral V proteins have the potential to inhibit all of them.Innate immunity is stimulated by viruses in part via RNA. dsRNA 2 specifically is present in several forms: viral genomes, single-stranded RNA virus replication intermediates, DNA virus symmetric transcription products, defective viral particles, and debris from lysed cells (1). Although extracellular dsRNA is sensed by Toll-like receptor 3 (TLR3), intracellular dsRNA is detected in part by the RNA helicases retinoic acidinducible gene 1 (RIG-I) and melanoma differentiation-associated gene 5 (Mda-5). These receptors signal through Toll-IL-1R (TIR) domain containing adaptor-inducing IFN␤ (TRIF) and IFN␤ promoter stimulator 1 (IPS1), respectively, to activate the kinases TANK-binding kinase 1 (TBK1) and inhibitor of B kinase ⑀ (IKKe). They, in turn, phosphorylate IFN regulatory factor 3 (IRF3), promoting its nuclear translocation and subsequent IFN-stimulated regulatory element-mediated transcription of IFN-stimulated genes (ISG), such as ISG56, as well as IFN and other cytokines. IFN then, through Janus kinase (JAK)/ STAT activation of IRF9, modulates microRNAs in addition to up-regulating itself and more ISGs to heighten the antiviral state and also to initiate the adaptive immune response by promoting dendritic cell maturation, memory T cell proliferation, and B cell differentiation (2-4).To control this immune response, pathogens and hosts have developed methods of down-regulation and evasion at a variety of different points (5-7). At the level of sensing infection, NS1 from influenza virus binds to and sequesters dsRNA and also interacts with RIG-I (5, 6, 8). At the level of signal transduction, the NS3/4A protease from hepatitis C virus cle...

Ventral furrow formation is a key morphogenetic event during Drosophila gastrulation that leads to the internalization of mesodermal precursors. While genetic analysis has revealed the genes involved in the specification of ventral furrow cells, few of the structural proteins that act as mediators of ventral cell behavior have been identified. A comparative proteomics approach employing difference gel electrophoresis was used to identify more than fifty proteins with altered abundance levels or isoform changes in ventralized versus lateralized embryos. Curiously, the majority of protein differences between these embryos appeared well before gastrulation, only a few protein changes coincided with gastrulation,suggesting that the ventral cells are primed for cell shape change. Three proteasome subunits were found to differ between ventralized and lateralized embryos. RNAi knockdown of these proteasome subunits and time-dependent difference-proteins caused ventral furrow defects, validating the role of these proteins in ventral furrow morphogenesis.

Mumps virus, like other paramyxoviruses in the Rubulavirus genus, encodes a V protein that can assemble a ubiquitin ligase complex from cellular components, leading to the destruction of cellular signal transducer and activator of transcription (STAT) proteins. While many V proteins target the interferon-activated STAT1 or STAT2 protein, mumps virus V protein is unique in its ability to also target STAT3 for ubiquitin modification and proteasome-mediated degradation. Here we report that a single amino acid substitution in the mumps virus V protein, E95D, results in defective STAT3 targeting while maintaining the ability to target STAT1. Results indicate that the E95D mutation disrupts the ability of the V protein to associate with STAT3. A recombinant mumps virus carrying the E95D mutation in its P and V proteins replicates normally in cultured cells but fails to induce targeting of STAT3. Infection with the recombinant virus results in the differential regulation of a number of cellular genes compared to wild-type mumps virus and increases cell death in infected cells, producing a large-plaque phenotype.

Ventral furrow formation is the first morphogenetic movement to occur during Drosophila gastrulation causing the internalization of mesodermal precursors. A previous proteomic screen for ventral-specific proteome changes identified a set of about forty "difference-proteins" that spanned many cellular functions. To understand the connections between these disparate proteins, we initiated a pathway-building scheme using cycles of protein expression manipulation and proteome analysis. This pathway-building exercise started with the proteasomal subunit, Pros35, one of three proteasome subunits found to be ventral-specific difference-proteins. Here we show that Pros35 is a key regulator in ventral furrow formation. Altering the level of Pros35 led to ventral furrow defects. Proteome analysis of the changes induced by Pros35 RNAi showed extensive overlap with the original set of ventral-specific difference-proteins. One of the most prominent changes was in the extracellular iron carrier, Transferrin (Tsf1). Tsf1 is normally less abundant in ventral cells relative to lateral cells; however, RNAi of Pros35 in ventralized embryos negated this ventral-specific difference. Increasing Tsf1 in wild-type embryos blocked ventral furrow formation and caused proteome changes that were similar to the previously seen ventral-specific difference-proteins, including Pros35, which indicates the existence of an unprecedented regulatory loop between the proteasome and iron homeostasis. Additionally, we show that the iron regulatory protein, Irp-1A, also plays an important role in ventral furrow formation. Together these three proteins are part of a regulatory loop that coordinately controls a large number of ventral-specific protein changes.

Due to rise in number of vehicles the traffic management has become a major problem. Manual traffic system is not efficient. This paper presents adaptive traffic management system using Internet of Things (IoT) and Image processing. The proposed system has capability to analyze real time data using image processing. Using cameras, different lanes are monitored constantly. The data obtained from different lanes are examined. Detection and counting of number of vehicles in each lane is done by using image processing. The count from each lane is sent to the central processing unit. According to the count of vehicles algorithm calculates waiting time for each lane, then the signal lights will be decided. This system reduces the average waiting time and increases the efficiency of traffic clearance. The system also reduces the pollution due CO2 emission and useful in emergency situations, thus being adaptive traffic management using Internet of Things (IoT).