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Abstract Senescence, a hallmark of aging, is a factor in age-related diseases (ARDs). Therefore, targeting senescence is widely regarded as a practicable method for modulating the effects of aging and ARDs. Here, we report the identification of regorafenib, an inhibitor of multiple receptor tyrosine kinases, as a senescence-attenuating drug. We identified regorafenib by screening an FDA-approved drug library. Treatment with regorafenib at a sublethal dose resulted in effective attenuation of the phenotypes of βPIX knockdown- and doxorubicin-induced senescence and replicative senescence in IMR-90 cells; cell cycle arrest, and increased SA-β-Gal staining and senescence-associated secretory phenotypes, particularly increasing the secretion of interleukin 6 (IL-6) and IL-8. Consistent with this result, slower progression of βPIX depletion-induced senescence was observed in the lungs of mice after treatment with regorafenib. Mechanistically, the results of proteomics analysis in diverse types of senescence indicated that growth differentiation factor 15 and plasminogen activator inhibitor-1 are shared targets of regorafenib. Analysis of arrays for phospho-receptors and kinases identified several receptor tyrosine kinases, including platelet-derived growth factor receptor α and discoidin domain receptor 2, as additional targets of regorafenib and revealed AKT/mTOR, ERK/RSK, and JAK/STAT3 signaling as the major effector pathways. Finally, treatment with regorafenib resulted in attenuation of senescence and amelioration of porcine pancreatic elastase-induced emphysema in mice. Based on these results, regorafenib can be defined as a novel senomorphic drug, suggesting its therapeutic potential in pulmonary emphysema.

The South Korean aquaculture industry has incurred considerable production losses due to various infectious diseases. Artificially synthesized polyinosinic–polycytidylic acid (poly I:C), structurally similar to double-stranded RNA (dsRNA) and cytidine–phosphate–guanosine oligodeoxynucleotides, can enhance immune responses and protect against diseases. Here, we investigated dsRNA molecules with different cytidine–phosphate–guanosine (CpG) motifs (dsRNA-CpGMix) as fused agents to treat Miamiensis avidus and viral hemorrhagic septicemia virus (VHSV) infection in olive flounders. We further investigated the efficacy of specific sequence motifs in dsRNA in modulating immunostimulatory effects. Fish treated with poly I:C or dsRNA-CpGMix exhibited higher survival rates than the control group. Olive flounder leukocytes stimulated with poly I:C or dsRNA-CpGMix showed increased scuticocidal activity in the presence of inactivated immune sera. dsRNA with CpG motif sequences induced higher resistance against M. avidus and VHSV infections than dsRNA without CpG motif sequences, and the dsRNA-CpGMix group showed upregulated ISG15 or Mx compared to the dsRNA-GFP group. Thus, dsRNA containing CpG motifs can be used as effective immunostimulants to enhance resistance against viral and parasitic diseases in olive flounder. The specific sequences of the CpG motifs in dsRNA may be important for enhancing immune responses and resistance against M. avidus and VHSV infections in olive flounders.

Abstract Aggregation of misfolded alpha-synuclein (α-synuclein) is a central player in the pathogenesis of neurodegenerative diseases. Therefore, the regulatory mechanism underlying α-synuclein aggregation has been intensively studied in Parkinson’s disease (PD) but remains poorly understood. Here, we report p21-activated kinase 4 (PAK4) as a key regulator of α-synuclein aggregation. Immunohistochemical analysis of human PD brain tissues revealed an inverse correlation between PAK4 activity and α-synuclein aggregation. To investigate their causal relationship, we performed loss-of-function and gain-of-function studies using conditional PAK4 depletion in nigral dopaminergic neurons and the introduction of lentivirus expressing a constitutively active form of PAK4 (caPAK4; PAK4S445N/S474E), respectively. For therapeutic relevance in the latter setup, we injected lentivirus into the striatum following the development of motor impairment and analyzed the effects 6 weeks later. In the loss-of-function study, Cre-driven PAK4 depletion in dopaminergic neurons enhanced α-synuclein aggregation, intracytoplasmic Lewy body-like inclusions and Lewy-like neurites, and reduced dopamine levels in PAK4DAT-CreER mice compared to controls. Conversely, caPAK4 reduced α-synuclein aggregation, as assessed by a marked decrease in both proteinase K-resistant and Triton X100-insoluble forms of α-synuclein in the AAV-α-synuclein-induced PD model. Mechanistically, PAK4 specifically interacted with the NEDD4-1 E3 ligase, whose pharmacological inhibition and knockdown suppressed the PAK4-mediated downregulation of α-synuclein. Collectively, these results provide new insights into the pathogenesis of PD and suggest PAK4-based gene therapy as a potential disease-modifying therapy in PD.

Gene expression: The role of a regulatory enzyme in disease An enzyme that regulates an important controller of gene expression may offer a therapeutic target for cancer and other diseases. cAMP response element-binding protein (CREB) interacts with various other proteins to switch a myriad of target genes on and off in different cells. A review by Eung-Gook Kim, Eun-Young Shin and colleagues at Chungbuk National University, Cheongju, South Korea, explores the interplay between CREB and an enzyme called p21-activated kinase 4 (PAK4) in human health and disease. PAK4, for example, has been shown to promote CREB’s gene-activating function in prostate cancer, and PAK4 overexpression is a feature of numerous other tumor types. Disruptions in PAK4-mediated regulation of CREB activity have also been observed in neurons affected by Parkinson’s disease. The authors see strong clinical promise in further exploring the biology of the PAK4-CREB pathway.

Objective: Miniscrew-assisted maxillary expansion devices are frequently used for patients with calcified midpalatal sutures. This study aimed to evaluate the effects of two bone-borne maxillary expansion appliances on the cranial and circummaxillary sutures by comparing cone-beam computed tomography (CBCT) images before and after transverse maxillary expansion. Methods: A total of 81 patients (women = 58, men = 23) were treated with either a C-expander (n = 44) or an ATOZ expander (n = 37). CBCT images were obtained before (T0) and after (T1) maxillary expansion, and the widths of 10 circummaxillary sutures were measured in the sagittal, coronal, and axial planes. The Wilcoxon signed-rank test was used to compare the changes in suture width between the C-expander and ATOZ groups, and statistical significance was set at P < 0.05. Results: The frontonasal, frontomaxillary, pterygomaxillary, nasomaxillary, internasal, intermaxillary, and midpalatal suture widths increased significantly after maxillary expansion in both the ATOZ and C-expander groups (both P < 0.05). The frontozygomatic, zygomaticomaxillary, and temporozygomatic suture widths decreased in the C-expander group (P < 0.05), whereas the frontozygomatic suture width increased significantly in the ATOZ group (P < 0.05). The width changes of the frontozygomatic, zygomaticomaxillary, temporozygomatic, pterygomaxillary, internasal, intermaxillary, and midpalatal sutures differed significantly between the two groups (P < 0.05). Conclusions: Both the C- and ATOZ expanders affected the suture width in the naso-maxillozygomatic region. The C-expander decreased the circum-zygomatic suture widths, whereas the ATOZ expander widened the frontozygomatic suture with no effect on other circummaxillary sutures. [ABSTRACT FROM AUTHOR]

Objective: To correlate temporomandibular joint (TMJ) morphology and position with cone-beam computed tomography (CBCT) images, Joint Vibration Analysis (JVA), and Jaw Tracker (JT) to develop a radiation-free, dynamic method for screening and monitoring the TMJ in orthodontic patients. Methods: A total of 236 orthodontic patients without symptoms of TMJ disorders who had undergone CBCT were selected for the JVA and JT tests in this cross-sectional study. TMJ position and morphology were measured using a three-dimensional analysis software. JT measurements involved six opening-closing cycles, and JVA measurements were performed using a metronome to guide the mouth opening-closing movements of the patients. The correlations among the three measuring devices were evaluated. Results: Abnormalities in condylar surface morphology affected the mandibular range of motion. The cut-off value results show that when various measurement groups are within a certain range, abnormalities may be observed in morphology (area under the curve, 0.81; P < 0.001). A 300/< 300 Hz ratio = 0.09 suggested abnormal morphology (P < 0.05). Correlations were observed among the maximum opening velocity, maximum vertical opening position, and joint spaces in the JT measurements. Correlations were also observed between the > 300/< 300 Hz ratio, median frequency, total integral, integral < 300 Hz, and peak frequency with joint spaces in the JVA measurements. Conclusions: JT and JVA may serve as rapid, non-invasive, and radiation-free dynamic diagnostic tools for monitoring and screening TMJ abnormalities before and during orthodontic treatment. [ABSTRACT FROM AUTHOR]

Highly textured (011)[100] Goss-oriented rolled sheet Fe-Al alloy is a promising magnetostrictive material for use in bending mode sensors and vibrational energy harvesters. In this paper, we performed magnetic field annealing (FA) to induce magnetic anisotropy in strips of highly textured Fe-Al. Prior work suggests FA as a viable alternative to stress annealing (SA), which leads to buckling of Fe-Al rolled sheet samples. The Fe-Al strips studied here exhibited tetragonal magnetostriction values ((3/2)λsat = λ∥ − λ⟂) of ∼136 ppm along their length, which corresponds to ∼78% of the single crystal value along a orientation. The effectiveness of FA on magnetic moment rotation was inferred by comparing post-FA magnetostriction measurements with the maximum possible yield, where λ∥= 0 and λ⟂= (3/2)λsat. Strain gauge data from the middle of the strip indicates that FA achieved ∼27% of the desired built-in uniaxial anisotropy along the parallel direction of the strip length, decreasing λ∥ by 25 ppm of the 95 ppm. Hall effect sensor data was used to assess the potential effect of FA on sensing and energy harvesting performance. FA improved the bending-stress-induced changes in magnetization near the clamped end of the strips by ∼45%. These results suggest that the FA was more effective in the region near the end of the strip than toward the middle of the strip, which we explain may be a result of the use of high temperature permanent magnets at the ends of the strips for the FA protocol.

Magnetostrictive Fe-Ga and Fe-Al alloys are promising materials for use in bending-mode vibrational energy harvesters. For this study, 50.8 mm × 5.0 mm × 0.5 mm strips of Fe-Ga and Fe-Al were cut from 0.50-mm thick rolled sheet. An atmospheric anneal was used to develop a Goss texture through an abnormal grain growth process. The anneal lead to large (011) grains that covered over 90% of sample surface area. The resulting highly-textured Fe-Ga and Fe-Al strips exhibited saturation magnetostriction values (λsat = λ∥ − λ⊥) of ∼280 ppm and ∼130 ppm, respectively. To maximize 90° rotation of magnetic moments during bending of the strips, we employed compressive stress annealing (SA). Samples were heated to 500°C, and a 100-150 MPa compressive stress was applied while at 500°C for 30 minutes and while being cooled. The effectiveness of the SA on magnetic moment rotation was inferred by comparing post-SA magnetostriction with the maximum possible yield of rotated magnetic moments, which is achieved when λ∥ = λsat and λ⊥ = 0. The uniformity of the SA along the sample length and the impact of the SA on sensing/energy harvesting performance were then assessed by comparing pre- and post-SA bending-stress-induced changes in magnetization at five different locations along the samples. The SA process with a 150 MPa compressive load improved Fe-Ga actuation along the sample length from 170 to 225 ppm (from ∼60% to within ∼80% of λsat). The corresponding sensing/energy harvesting performance improved by as much as a factor of eight in the best sample, however the improvement was not at all uniform along the sample length. The SA process with a 100 MPa compressive load improved Fe-Al actuation along the sample length from 60 to 73 ppm (from ∼46% to ∼56% of λsat, indicating only a marginally effective SA and suggesting the need for modification of the SA protocol. In spite of this, the SA was effective at improving the sensing/energy harvesting performance by a factor of ∼2.5 in the best sample. As with the Fe-Ga strip, improvement in performance was quite varied along the strip length.

The magnetic force microscopy technique is employed to explain the various magnetic properties of the multilayer magnetostrictive iron-gallium nanowires. The different effects of the aspect ratio on the anisotropy effects of the nanowires are also discussed.

We have investigated a method for measuring the dimensions of an individual multilayered Fe- Ga/Cu nanowire (NW) as it changes with induced magnetization. In this study, we demonstrate the proposed approach and establish this as a viable method for measuring the magnetostrictive behavior of an individual Fe-Ga/Cu NW using atomic force microscopy (AFM). When an external magnetic field (~300 Oe) was applied perpendicular to the NW axis, the NW length appeared minimized. When a field (~1000 Oe) was applied parallel to the NW axis, the height profile of the NW was found to be higher than in the case with no parallel external field. Since both ends of the NW were welded to the substrate, the magnetic field induced dimensional change of the NW caused deflection of the NW in the upward direction, which was significant enough to be detected by AFM. An average height difference of 15nm was measured with and without an applied field which was then used to calculate the magnetostriction of the multilayered NW. [ABSTRACT FROM AUTHOR]

As one of the most attractive large-capacity energy storage systems, vanadium redox flow batteries have many advantages such as a rapid response, long service life, environment-friendliness, and non-flammability. However, the high cost of the vanadium electrolyte has hindered the broad market penetration. The price of the vanadium electrolyte can be significantly reduced if a cost-competitive technology for producing the electrolyte from recycled V2O5 may be used. Accordingly, the effects of the impurities in the recycled V2O5 on the performance of vanadium redox flow batteries (VRFBs) must be understood. However, there have been very few published studies on the effects of these electrolyte impurities. In order to establish a reliable electrolyte specification for VRFB developers, we analyzed the influence of a wide range of potential metal impurities or additives on the electrochemical activity, diffusion of vanadium ions, and battery performance of these systems. The investigated impurities include Li+, Na+, K+, Mg2+, Cr3+, Mn2+, Fe2+/F3+, Co2+, Ni2+, Cu2+, Zn2+, andMo6+ ions.Most of the metal ions affected the V2+/V3+ redox reaction rather than the VO2+/VO2 + redox reaction. On the basis of our findings, we suggest detailed specifications that may be used to manage the quality of the vanadium electrolyte [ABSTRACT FROM AUTHOR]

Mitsugumin 53 (MG53) is an E3 ligase that interacts with and ubiquitinates insulin receptor substrate-1 (IRS-1) in skeletal muscle; thus, an MG53-IRS-1 interaction disruptor (MID), which potentially sensitizes insulin signaling with an elevated level of IRS-1 in skeletal muscle, is an excellent candidate for treating insulin resistance. To screen for an MID, we developed a bimolecular luminescence complementation system using an N-terminal luciferase fragment fused with IRS-1 and a C-terminal luciferase fragment fused with an MG53 C14A mutant that binds to IRS-1 but does not have E3 ligase activity. An MID, which was discovered using the bimolecular luminescence complementation system, disrupted the molecular association of MG53 with IRS-1, thus abolishing MG53-mediated IRS-1 ubiquitination and degradation. Thus, the MID sensitized insulin signaling and increased insulin-elicited glucose uptake with an elevated level of IRS-1 in C2C12 myotubes. These data indicate that this MID holds promise as a drug candidate for treating insulin resistance. [ABSTRACT FROM AUTHOR]

Aberrant sialylation has long been correlated with human cancer. Increased ST6 Gal I (β-galactoside α 2, 6 sialyltransferase) and consequently higher levels of cell-surface α 2, 6 sialylation has been associated with human colorectal cancer (CRC) metastasis. We have extensive circumstantial data that sialylation is connected to cancer metastasis, but we do not understand in detail how sialylation can switch on/off multiple steps in cancer metastasis. To investigate the molecular mechanism underlying the ST6Gal I-mediated metastasis of CRC, we silenced the ST6Gal I gene in a metastatic SW620 CRC cell line (SW620-shST6Gal I) and examined the metastatic behavior of the cells. We found that various hallmarks of metastatic ability were considerably enhanced in ST6Gal 1-depleted SW620 clones, as assessed both in vitro and in vivo. In particular, the metastasis suppressor, KAI1, was down-regulated in ST6Gal I-deficient SW620 clones. This reflected the increased exosome-mediated exportation of KAI1, and was associated with a decrease in the KAI1-mediated inhibition of integrin. These findings indicate that gene silencing of ST6Gal I could enhance metastasis of CRC by down-regulating KAI1 activity and rescuing its negative effects on integrin signaling. [ABSTRACT FROM AUTHOR]

Abnormal glycosylation due to dysregulated glycosyltransferases and glycosidases is a key phenomenon of many malignancies, including colorectal cancer (CRC). In particular, increased ST6 Gal I (b-galactoside a 2, 6 sialyltransferase) and subsequently elevated levels of cell-surface a 2, 6-linked sialic acids have been associated with metastasis and therapeutic failure in CRC. As many CRC patients experience metastasis to the liver or lung and fail to respond to curative therapies, intensive research efforts have sought to identify the molecular changes underlying CRC metastasis. ST6 Gal I has been shown to facilitate CRC metastasis, and we believe that additional investigations into the involvement of ST6 Gal I in CRC could facilitate the development of new diagnostic and therapeutic targets. This review summarizes how ST6 Gal I has been implicated in the altered expression of sialylated glycoproteins, which have been linked to CRC metastasis, radioresistance, and chemoresistance. [ABSTRACT FROM AUTHOR]

We demonstrated that white emission can be obtained from nano-structured top-emitting organic light-emitting diodes (TEOLEDs) based on a blue emitting layer (EML). The nano-structured TEOLEDs were fabricated on nano-patterned substrates, in which both optical micro-cavity and scattering effects occur simultaneously. Due to the combination of these two effects, the electroluminescence spectra of the nano-structured device with a blue EML exhibited not only blue but also yellow colours, which corresponded to the intrinsic emission of the EML and the resonant emission of the micro-cavity effect. Consequently, it was possible to produce white emission from nano-structured TEOLEDs without employing a multimode micro-cavity. The intrinsic emission wavelength can be varied by altering the dopant used for the EML. Furthermore, the emissive characteristics turned out to be strongly dependent on the nano-pattern sizes of the nano-structured devices. [ABSTRACT FROM AUTHOR]

Rural landscapes in many parts of the world no longer exist solely for agricultural production and therefore face change to accommodate various socio-economic demands. Hence, there is a need to improve our understanding of public attitudes—in particular, their openness—toward such change. This article discusses predictors of openness to landscape change in rural areas based on the results of an online survey conducted to develop attitudinal measurement scales for landscape change and rural landscape in England. Landscape change in rural areas was not welcomed by the respondents mainly due to their high level of affirmative attitudes to rural landscapes. However, the study suggests that there is a possibility, especially among younger cohorts, that increasing levels of education will lead to greater openness to landscape change. [ABSTRACT FROM PUBLISHER]

Abstract  A novel glucose sensor was constructed, and its analytical potential examined. A chip-type three-electrode system for use in a flow-type electrochemical glucose sensor was fabricated using a UV lithography technique on a glass slide. An Ag/AgCl reference electrode was made by electroplating silver onto a Pt electrode and dipping in a saturated KCl solution for 30 min. In addition, a glucose-sensing electrode was fabricated using a two-photon adsorbed photopolymerization technique with a photo-reactive resin containing a glucose oxidase enzyme, ferrocene mediator, non-ionic surfactant, and carbon nanotubes. The cyclic voltammetry of the potassium ferrocyanide in the Pt sensor system showed a stable electrode condition. The response of the modified Pt sensor confirms the feasibility of using a two-photon adsorbed photopolymerization technique for the easy fabrication of functional biosensors. [ABSTRACT FROM AUTHOR]

We investigate the utility of Langmuir adsorption measurements for characterizing nanoparticle−substrate interactions. Spherical CdSe/ZnS core−shell nanoparticles were chosen as representative particles because of their widespread use in biological labeling measurements and their relatively monodisperse dimensions. In particular, the quantum dots were functionalized with 11-mercaptoundecanoic acid, and we utilized an amine-terminated self-assembled monolayer (SAM) as a model substrate. SAMs with different end-groups (−CH3and −COOH) were also considered to contrast with the adsorption behavior on the amine-terminated SAM substrates. We followed the kinetics of nanoparticle adsorption on the aminosilane layer by quartz crystal microgravimetry (QCM) over a range of particle concentrations and determined the corresponding Langmuir adsorption isotherms. Analysis of both equilibrium adsorption and kinetic adsorption data allowed us to determine a consistent value of the Langmuir adsorption equilibrium constant for the amine-terminated SAM at room temperature (KL≈ 2.7 (μmol/L)−1), providing a useful characterization of the nanoparticle−substrate interaction. The effect of varying solution pH on Langmuir adsorption was also investigated in order to gain insight into the role of electrostatic interactions on nanoparticle adsorption. The equilibrium extent of adsorption was found to be maximum at about pH 7. These changes of nanoparticle adsorption were further quantified and validated by X-ray photoelectron spectroscopy (XPS) and confocal fluorescence microscopy measurements. We conclude that Langmuir adsorption measurements provide a promising approach for quantifying nanoparticle−substrate interactions. [ABSTRACT FROM AUTHOR]

This study aimed to analyze the treatment effects of miniscrew-assisted rapid palatal expansion using dental cone-beam computed tomography (CT) in adolescents. The study group comprised 20 patients who underwent cone-beam CT before and after miniscrew-assisted rapid palatal expansion (MARPE). Paired t-test and analysis of variance were used to analyze significant differences before and after treatment. Skeletal and dentoalveolar measurements indicated highly significant differences, with these expansions increasing progressively in the downward direction. The extent of anterior midpalatal suture expansion was significantly greater than that of the posterior part. Significant expansion was observed at the nasal width and inter-zygomaticomaxillary suture distance. MARPE can expand the craniofacial skeletal structure to the uppermost nasal bone and lateral to the zygomaticomaxillary sutures. The expansion was greater in the lower maxillary part, demonstrating an overall triangular maxillary expansion. The midpalatal suture opening was greater in the anterior than in the posterior side, whereas the posterior suture opening proportion was higher than that of the conventional rapid palatal expansion previously reported.

Background: We conducted a narrative review of studies analyzing the occlusal and mandibular plane angles after maxillary molar distalization using temporary skeletal anchorage devices (TADs). An original preliminary investigation was conducted on the occlusal and mandibular plane angle changes according to the design of TAD-supported distalization. Methods: We included 51 participants stratified into three groups (buccal TAD, lingual arch TAD, and pendulum TAD) who underwent lateral cephalography before and after treatment. The paired t-test and ANOVA were used to analyze the significant differences among the groups. Results: The pterygoid vertical to maxillary second molar (p < 0.01) significantly differed before and after treatment within each group (group 1, 1.29 ± 1.73 mm; group 2, 2.01 ± 1.46 mm; group 3, 1.12 ± 1.43 mm). The angle between the Frankfort horizontal and anatomical occlusal plane increased by 1.96 ± 2.88° (p < 0.05) and 2.51 ± 2.57° (p < 0.01) in groups 2 and 3, respectively. Conclusions: The measured variables did not significantly differ among the three groups. The Frankfort-mandibular plane angle and functional occlusal plane did not change after maxillary molar distalization. The anatomical occlusal plane is influenced by the anterior teeth and may be unrelated to maxillary molar distalization. Further studies are required to verify the exact relationship between maxillary molar distalization and occlusal plane angle.

The article discusses bioinformatic and functional data link integrin-mediated cell adhesion to cellular senescence. It mentions that role for aberrant integrin signaling in senescence, but how senescence alters cell-matrix adhesion and how this contributes to the senescent phenotype are poorly understood. It also mentions that cellular senescence is associated with cell cycle arrest, which can be due to either increased expression of the cyclin-dependent kinase inhibitors.

To address whether mitochondrial biogenesis is essential for skeletal myogenesis, C2C12 myogenesis was investigated after knockdown of NADH dehydrogenase (ubiquintone) flavoprotein 1 (NDUFV1), which is an oxidative phosphorylation complex I subunit that is the first subunit to accept electrons from NADH. The NDUFVI knockdown enhanced C2C12 myogenesis by decreasing the NAD+/NADH ratio and subsequently inactivating SIRT1 and SIRT1 activators (pyruvate, SRT1720, and resveratrol) abolished the NDUFV1 knockdown-induced myogenesis enhancement. However, the insulin-elicited activation of insulin receptor β (IRβ) and insulin receptor substrate-1 (IRS-1) was reduced with elevated levels of protein-tyrosine phosphatase 1B after NDUFV1 knockdown in C2C12 myotubes. The NDUFV1 knockdown-induced blockage of insulin signaling was released by protein-tyrosine phosphatase 1B knockdown in C2C12 myotubes, and we found that NDUFV1 or SIRT1 knockdown did not affect mitochondria biogenesis during C2C12 myogenesis. Based on these data, we can conclude that complex I dysfunction-induced SIRT1 inactivation leads to myogenesis enhancement but blocks insulin signaling without affecting mitochondria biogenesis. [ABSTRACT FROM AUTHOR]

Our functional mapping study of murine gammaherpesvirus 68 (MHV-68, or γ HV-68) revealed that a mutant harboring a transposon at the ORF49 locus (ORF49null) evidenced a highly attenuated in vitro growth. ORF49 resides adjacent to and in an opposite direction from RTA, the primary switch of the gammaherpesvirus life cycle. A FLAG-tagged ORF49 protein was able to transcomplement ORF49null, and a revertant of ORF49null restored its attenuated growth to a level comparable to that of the wild type. The FLAG-tagged ORF49 protein promoted the ability of RTA to activate downstream target promoters and enhanced virus replication from the ORF50null virus in the presence of RTA. Furthermore, ORF49 enhanced wild-type virus replication by increasing the RTA transcript levels. Our data indicate that ORF49 may perform an important function in MHV-68 replication in cooperation with RTA. [ABSTRACT FROM AUTHOR]

The human cytomegalovirus (HCMV) 72-kDa immediate-early 1 (IE1) protein is thought to modulate cellular antiviral functions impacting on promyelocytic leukemia (PML) nuclear bodies and signal transducer and activator of transcription (STAT) signaling. IE1 consists of four distinct regions: an amino-terminal region required for nuclear localization, a large central hydrophobic region responsible for PML targeting and transactivation activity, an acidic domain, and a carboxyl-terminal chromatin tethering domain. We found that the acidic domain of IE1 is required for binding to STAT2. A mutant HCMV encoding IE1Δ (421-475) with the acidic domain deleted was generated. In mutant virus-infected cells, IE1(Δ 421-475) failed to bind to STAT2. The growth of mutant virus was only slightly delayed at a high multiplicity of infection (MOI) but was severely impaired at a low MOI with low-level accumulation of viral proteins. When cells were pretreated with beta interferon, the mutant virus showed an additional 1,000-fold reduction in viral growth, even at a high MOI, compared to the wild type. The inhibition of STAT2 loading on the target promoter upon infection was markedly reduced with mutant virus. Furthermore, sumoylation of IE1 at this acidic domain was found to abolish the activity of IE1 to bind to STAT2 and repress the interferon-stimulated genes. Our results provide genetic evidence that IE1 binding to STAT2 requires the 55-amino-acid acidic domain and promotes viral growth by interfering with interferon signaling and demonstrate that this viral activity is negatively regulated by a cellular sumoylation pathway. [ABSTRACT FROM AUTHOR]