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SUMMARY: Proline metabolism plays a crucial role in both environmental stress responses and plant growth. However, the specific mechanism by which proline contributes to abiotic stress processes remains to be elucidated. In this study, we utilized atrzf1 (Arabidopsis thaliana ring zinc finger 1) as a parental line for T‐DNA tagging mutagenesis and identified a suppressor mutant of atrzf1, designated proline content alterative 31 (pca31). The pca31 mutant suppressed the insensitivity of atrzf1 to dehydration stress during early seedling growth. Using Thermal Asymmetric Interlaced‐PCR, we found that the T‐DNA of pca31 was inserted into the promoter region of the At2g22620 gene, which encodes the cell wall enzyme rhamnogalacturonan lyase 1 (RGL1). Enzymatic assays indicated that RGL1 exhibited rhamnogalacturonan lyase activity, influencing cell wall pectin composition. The decrease in RGL1 gene expression suppressed the transcriptomic perturbation of the atrzf1 mutant. Silencing of the RGL1 gene in atrzf1 resulted in a sensitive phenotype similar to pca31 under osmotic stress conditions. Treatment with mannitol, salt, hydrogen peroxide, and abscisic acid induced RGL1 expression. Furthermore, we uncovered that RGL1 plays a role in modulating root growth and vascular tissue development. Molecular, physiological, and genetic experiments revealed that the positive modulation of RGL1 during abiotic stress was linked to the AtRZF1 pathway. Taken together, these findings establish that pca31 acts as a suppressor of atrzf1 in abiotic stress responses through proline and cell wall metabolisms. Significance Statement: RGL1 exhibits rhamnogalacturonan lyase activity, which contributes to the modulation of pectin composition, vascular tissue development, and root growth. RGL1, as a suppressor of E3 ubiquitin ligase AtRZF1, functions as a positive modulator in proline synthesis and cell wall metabolism under osmotic stress conditions. [ABSTRACT FROM AUTHOR]

Noise from firearms is well known to be harmful to human hearing. This problem has been addressed by various military units through the use of muzzle suppressors. However, as suppressor technology has advanced, shooters report hearing the mechanical action of gas-operated semi-automatic rifles (ArmaLite Rifle Model 15 style aka AR-15) as being louder than the suppressed muzzle noise. This study aims to evaluate if harmful noise is present in the shooter's ear, even when impulse noise emanating from the muzzle is suppressed. To characterize the impulse noise of the firearm action caused by the reciprocation of the bolt carrier group (BCG) and subsequent impact when it returns to battery (the forward locked position), the muzzle of a rifle was placed through a constructed plywood wall, and the noise of the action/breech was measured independently from the muzzle noise. This research finds that the impact of the BCG returning to battery (132 dBZ) has the potential to be harmful to the shooter's hearing even when the noise from the muzzle is effectively suppressed. [ABSTRACT FROM AUTHOR]

Whilst research has assessed the multidimensionality of organizational stressors, severity has been overlooked. This study aimed to develop and validate a severity response scale, before examining the relationship between organizational stressor severity, physical health, and well-being, and if severity mediated the relationship between stressor occurrence and outcomes. 403 sport performers (186 male; Mage = 21.43) completed a questionnaire assessing study variables and data were analysed using structural equation modelling. Organizational stressor severity had a significant, negative relationship with physical health (r = -.374, p <.001), but notwith hedonic or eudaimonic well-being (p =.624 and.203). Direct effects were found for neuroticism (β = -0.632, p <.001) and stressor frequency (β = -0.226, p =.016) on physical health; however, when adding severity as a mediator, stressor frequency positively predicted stressor severity (β = 1.025, p <.001); however, the indirect effect of stressor frequency on physical health via stressor severity (β = -0.018, p =.885) and the direct effect between stressor frequency and physical health (β = -0.191, p =.204) were non-significant. These novel findings highlight that higher stressor severity is associated with higher physical health complaints, and that repeated exposure to stressors may heighten an vulnerability to future stressors. [ABSTRACT FROM AUTHOR]

The noise created when a firearm is fired has a lot of adverse effects on humans and the environment, so analyzing and attenuating this noise is essential. This study aimed to examine propellant flow and the sound generated by this flow using a hybrid computational fluid dynamics and computational aeroacoustics method. The compatibility of this numerical study was also validated by comparing it with the experimental result. The impact of critical parameters, such as the shape and number of baffles of the suppressor, was studied. Finally, the overpressure and acoustics results of different suppressors were compared with each other and with the unsuppressed condition. According to the result, the suppressor with curved baffles shows a better performance. When the exit pressure at the tip of suppressor compared to the initial inlet pressure to the suppressor the percentage drop in overpressure was 76.01%, 78.79% and 81.3% for the suppressor with one, three and five curved baffles, respectively. For condition without suppressor, 169.49 dB of SPL was recorded. When using a suppressor without a baffle, this value was reduced to 162.13 dB. For the suppressor with one, three and five curved baffles, the SPL value was 160.234 dB, 159.43 dB and 158.11 dB, respectively. Generally, this study shows that the suppressor's efficiency is directly proportional to its shape and number of baffles. [ABSTRACT FROM AUTHOR]

The regulation of intracellular pH in yeast (Saccharomyces cerevisiae) cells is critical for cell function and viability. In yeast, protons (H+) can be excreted from the cell by plasma membrane ATPase PMA1 and pumped into vacuoles by vacuolar H+-ATPase. Because PMA1 is critical to the survival of yeast cells, it is unknown whether other compensatory components are involved in pH homeostasis in the absence of PMA1. To elucidate how intracellular pH is regulated independently of PMA1, we employed a screening approach by exposing the yeast haploid deletion mutant library (ver 4.0) to the selective plant plasma membrane H+-ATPase inhibitor PS-1, which we previously reported. After repeated screenings and verification, we identified two proteins, Aly1 and Aly2, that play a role in the regulation of intracellular pH when PMA1 is deficient. Our research uncovers a new perspective on the regulation of intracellular pH related to PMA1 and also preliminarily reveals a role for Aly1 and Aly2 in the regulation of intracellular pH.

Introduction: Citrus chlorotic dwarf-associated virus (CCDaV) is an economically important citrus virus associated with leaf curling, deformation, and chlorosis found in China. Plants have evolved RNA silencing to defend against viral infections; however, the mechanism by which CCDaV suppresses RNA silencing in citrus remains unknown. Methods: Six proteins encoded by CCDaV were ectopically expressed in Nicotiana benthamiana 16c using the pCHF3 vector to identify RNA-silencing suppression activities. Results: V2 protein encoded by CCDaV suppressed local RNA silencing and systemic RNA silencing triggered by GFP RNA, but did not impede short-distance movement of the RNA silencing signal in N. benthamiana 16c. GFP fluorescence observations showed that the ability of V2 protein to suppress RNA silencing was weaker than tomato bushy stunt virus P19. Deletion analysis showed that the putative nuclear localization signal (NLS, 25-54 aa) was involved in the RNA silencing suppression activity of V2 protein. Furthermore, V2 protein cannot block dsRNA-triggered RNA silencing. The subcellular localization assay suggested that V2 protein was localized to nucleus of N. benthamiana. Conclusion: Overall, the results of this study demonstrate that CCDaV-V2 acts as an activity of silencing suppression. This is the first reported RNA-silencing suppressor encoded by Citlodavirus and will be valuable in revealing the molecular mechanism of CCDaV infection. [ABSTRACT FROM AUTHOR]

A powerful soundwave from shooting a firearm can travel great distances harming the shooter as well as anyone or anything standing nearby. To address this, a nontypical firearm suppressor is used. A suppressor muffles the soundwave to a level that is usually safe for the shooter and its environment. A suppressor usually has 3-4 main parts that highly influence its effectiveness. The study aims to investigate how non-typical baffle design and placement selection influence the suppressor effectiveness. The study consists of 162 different non-typical baffle design configurations. Suppressors with 3 to 9 baffles, angled from 30 to 60 degrees as well as rounded from 10 mm to 30 mm radius, are simulated in a CFD environment. Suppressors with the best designs were then produced and evaluated on a firing range. At the firing range, a result of 32.24 dB delta was achieved and was only 4,67% different than the result presented in the CFD study, which was 30.8 dB, respectively. The 9 straight geometry baffle, angled at 30 degrees, suppressor, with an average chamber volume proved to be the most efficient and combined with a large initial chamber was the most efficient design in this study. [ABSTRACT FROM AUTHOR]

This study numerically examined the propellant flow from gunfire using the kω-SST turbulence model and their sound attenuation using the Ffowcs-Williams and Hawkins equations (FW-H). For simulation, a pressure-based solver and 3D axisymmetric geometry were used. The second-order implicit time approach and the second-order upwind scheme spatial discretization were used in the simulation. The maximum exit pressure was 3.748 MPa for the suppressor with a length of 70 mm and diameter of 20 mm. However, when the diameter suppressor increased by 1/6, the maximum exit pressure was reduced to 3.4961 MPa. When the length increased by 1/6, the maximum pressure became 3.3636 MPa. Lastly, when the diameter and length were increased by 1/6, the maximum exit pressure became 3.177 MPa. For this suppressor, 20.835 dB (12.29%) sound pressure level attenuation was achieved with 16.823 MPa (84.115%) overpressure reduction and 484.86 K or 32.32% temperature reduction. Generally, the attenuation increased with the increase in the suppressor's internal volume. [ABSTRACT FROM AUTHOR]

A firearm generates complex phenomena in muzzle flow and modelling the flow field around the projectile has great importance on high-intensity noise prediction. The negative effects of noise can be reduced using a suppressor which can be internally or externally attached to the barrel of a firearm. The purpose of this paper is to numerically and experimentally investigate the effect of the number and distance of circular ring baffles in the suppressor on the flow field and far field noise levels. Calculations were carried out in two-dimensional, axisymmetric, transient conditions and Ffowcs Williams and Hawkings acoustic analogy (FW-H) equations were solved to predict the far field noise. Nine cases including a gun without a suppressor, a suppressor without baffles, one, three, and five baffles which were placed at 20 mm intervals from the suppressor inlet, and one, three, five, seven, and nine baffles which were placed with equal intervals in the suppressor were simulated and compared; generations of noise during the shooting process were analyzed. The results showed that in the case without a suppressor, the peak sound pressure level was 156.1 dB at a 2.5 m distance, while this value decreased by nearly 7.6% in the case of the suppressor with seven baffles which has an average value of 144.2 dB. The results obtained here may provide a beneficial reference for predicting the muzzle noise and optimizing the number of baffles in the suppressor for small caliber gun systems. [ABSTRACT FROM AUTHOR]

Background: PIWI-interacting RNAs (piRNAs) are a class of noncoding RNAs originally reported in the reproductive system of mammals and later found to be aberrantly expressed in tumors. However, the function and mechanism of piRNAs in testicular cancer are not very clear. Methods: The expression level and distribution of piR-36249 were detected by RT-qPCR and immunofluorescence staining assay. Testicular cancer cell (NT2) progression was measured by CCK8 assay, colony formation assay and wound healing assay. Cell apoptosis was assessed by flow cytometry and western blot. RNA sequencing and dual-luciferase reporter assay were conducted to identify the potential targets of piR-36249. The relationship between piR-36249 and OAS2 or DHX36 was confirmed using overexpression assay, knockdown assay, pull-down assay and RIP assay. Results: piR-36249 is significantly downregulated in testicular cancer tissues compared to tumor-adjacent tissues. Functional studies demonstrate that piR-36249 inhibits testicular cancer cell proliferation, migration and activates the cell apoptosis pathway. Mechanically, we identify that piR-36249 binds to the 3′UTR of 2′-5′-oligoadenylate synthetase 2 (OAS2) mRNA. OAS2 has been shown in the literature to be a tumor suppressor modulating the occurrence and development of some tumors. Here, we show that OAS2 knockdown also promotes testicular cancer cell proliferation and migration. Furthermore, piR-36249 interacts with DHX36, which has been reported to promote translation. DHX36 can also bind to OAS2 mRNA, and knockdown of DHX36 increases OAS2 mRNA but downregulates its protein, indicating the enhancing effect of DHX36 on OAS2 protein expression. Conclusion: All these data suggest that piR-36249, together with DHX36, functions in inhibiting the malignant phenotype of testicular cancer cells by upregulating OAS2 protein and that piR-36249 may be used as a suppressor factor to regulate the development of testicular cancer.

Copyright of Journal of Ordnance Equipment Engineering is the property of Chongqing University of Technology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

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Copyright of Mycosystema is the property of Mycosystema Editorial Board and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

In the work, a theoretical analysis of high-voltage surge (HVS) passing from a power supply network through the path "voltage limiter - low-pass filter" is carried out, with the aim to evaluate the effectiveness of the filter on reducing the amplitude of the residual HVS at the input of electrical and/or radio-electronic equipment. As a low-pass filter (LPF), an L-shaped filter was chosen. Typical means of limiting the amplitude of the HVS (gas-filled arresters, varistors, and suppressors) are analyzed, and their shortcomings are shown. The justification of the equivalent scheme for theoretical analysis is given. The analysis revealed a direct relationship between the cut-off frequency of the LPF, which is determined by the nominal values of its elements, and the amplitude of the residual HVS at the mains input of the equipment, and also showed the effectiveness of inserting the LPF between the output of a typical HVS voltage limiter and the mains input of the mentioned equipment from the point of view of additional reduction of the residual HVS amplitude. References 18, figures 6. [ABSTRACT FROM AUTHOR]

Simple Summary: Gliomas are the most common primary brain tumors and are associated with significant mortality and morbidity. Less than 5% of patients with glioblastoma, the most common glioma histology, survive longer than five years. Therefore, searching for new biomarkers/molecular targets remains a constant issue in glioma research. One such target may be the metastasis suppressor BRMS1. BRMS1 interacts with critical steps of the metastatic cascade in many cancer entities. However, due to the low incidence of extra-cerebral glioma metastasis, the role of metastasis-associated proteins in gliomas remains poorly investigated. Still, the changes in behavior modulated by BRMS1 across different entities (e.g., affecting invasion, migration, and apoptosis) closely resemble the changes seen in gliomas. Additionally, BRMS1's interaction partners are commonly dysregulated in gliomas. Therefore, BRMS1 shows potential as a regulator of glioma behavior, and we present the first insights into BRMS1 expression in gliomas as a starting point for further investigations. The metastatic suppressor BRMS1 interacts with critical steps of the metastatic cascade in many cancer entities. As gliomas rarely metastasize, BRMS1 has mainly been neglected in glioma research. However, its interaction partners, such as NFκB, VEGF, or MMPs, are old acquaintances in neurooncology. The steps regulated by BRMS1, such as invasion, migration, and apoptosis, are commonly dysregulated in gliomas. Therefore, BRMS1 shows potential as a regulator of glioma behavior. By bioinformatic analysis, in addition to our cohort of 118 specimens, we determined BRMS1 mRNA and protein expression as well as its correlation with the clinical course in astrocytomas IDH mutant, CNS WHO grade 2/3, and glioblastoma IDH wild-type, CNS WHO grade 4. Interestingly, we found BRMS1 protein expression to be significantly decreased in the aforementioned gliomas, while BRMS1 mRNA appeared to be overexpressed throughout. This dysregulation was independent of patients' characteristics or survival. The protein and mRNA expression differences cannot be finally explained at this stage. However, they suggest a post-transcriptional dysregulation that has been previously described in other cancer entities. Our analyses present the first data on BRMS1 expression in gliomas that can provide a starting point for further investigations. [ABSTRACT FROM AUTHOR]

This review summarizes the progress in suppressor over the period from 1981 to 2022, with an emphasis on the last ten years. Although quite well investigated for decades, suppression techniques, along with their crucial roles in emerging analysis areas, have presented significantly scientific challenges. Apart from the intrinsic characters of vital components used, the design of system and the suppression mechanism are extremely important to improve the performance of suppressor for ion chromatography. Early packed-column suppressor must be regenerated off-line using an acid or base solution and had too large delay volume to possess sufficient suppression capacity. The trend is shifting toward the development of convenient, sustainable and high-performance suppressors in several ways: (i) the substantial improvement in the electrolytically or chemically regenerated suppression techniques; (ii) the miniaturization of conventional suppressors and the use of innovative designs to reduce the void volume and the overall band dispersion; and (iii) the application of ion reflux to suppressor for reducing the generation of waste. The topical review mainly focuses on the development and special applications of suppressors, spacing from the design and performance of suppressors, buffer generation, analyte enrichment and matrix isolation. Furthermore, the current challenges and future prospects regarding suppressors are discussed. [ABSTRACT FROM AUTHOR]

Daytime warm temperature elicits thermomorphogenesis in Arabidopsis by stabilizing the central thermoregulator PHYTOCHROME INTERACTING transcription FACTOR 4 (PIF4), whose degradation is otherwise promoted by the photoreceptor and thermosensor phytochrome B. PIF4 stabilization in the light requires a transcriptional activator, HEMERA (HMR), and is abrogated when HMR's transactivation activity is impaired in hmr-22. Here, we report the identification of a hmr-22 suppressor mutant, rcb-101, which surprisingly carries an A275V mutation in REGULATOR OF CHLOROPLAST BIOGENESIS (RCB). rcb-101/hmr-22 restores thermoresponsive PIF4 accumulation and reverts the defects of hmr-22 in chloroplast biogenesis and photomorphogenesis. Strikingly, similar to hmr, the null rcb-10 mutant impedes PIF4 accumulation and thereby loses the warm-temperature response. rcb-101 rescues hmr-22 in an allele-specific manner. Consistently, RCB interacts directly with HMR. Together, these results unveil RCB as a novel temperature signaling component that functions collaboratively with HMR to initiate thermomorphogenesis by selectively stabilizing PIF4 in the daytime.

ABSTRACT The Enterococcus faecalis acyl-acyl carrier protein (ACP) phosphate acyltransferase PlsX plays an important role in phospholipid synthesis and exogenous fatty acid incorporation. Loss of plsX almost completely blocks growth by decreasing de novo phospholipid synthesis, which leads to abnormally long-chain acyl chains in the cell membrane phospholipids. The ∆plsX strain failed to grow without supplementation with an appropriate exogenous fatty acid. Introduction of a ∆fabT mutation into the ∆plsX strain to increase fatty acid synthesis allowed very weak growth. The ∆plsX strain accumulated suppressor mutants. One of these encoded a truncated β-ketoacyl-ACP synthase II (FabO) which restored normal growth and restored de novo phospholipid acyl chain synthesis by increasing saturated acyl-ACP synthesis. Saturated acyl-ACPs are cleaved by a thioesterase to provide free fatty acids for conversion to acyl-phosphates by the FakAB system. The acyl-phosphates are incorporated into position sn1 of the phospholipids by PlsY. We report the tesE gene encodes a thioesterase that can provide free fatty acids. However, we were unable to delete the chromosomal tesE gene to confirm that it is the responsible enzyme. TesE readily cleaves unsaturated acyl-ACPs, whereas saturated acyl-ACPs are cleaved much more slowly. Overexpression of an E. faecalis enoyl-ACP reductase either FabK or FabI which results in high levels of saturated fatty acid synthesis also restored the growth of the ∆plsX strain. The ∆plsX strain grew faster in the presence of palmitic acid than in the presence of oleic acid with improvement in phospholipid acyl chain synthesis. Positional analysis of the acyl chain distribution in the phospholipids showed that saturated acyl chains dominate the sn1-position indicating a preference for saturated fatty acids at this position. High-level production of saturated acyl-ACPs is required to offset the marked preference of the TesE thioesterase for unsaturated acyl-ACPs and allow the initiation of phospholipid synthesis.

ABSTRACT It is uncertain whether PA1610|fabA is essential or dispensable for growth on LB-agar plates under aerobic conditions in Pseudomonas aeruginosa PAO1. To examine its essentiality, we disrupted fabA in the presence of a native promoter-controlled complementary copy on ts-plasmid. In this analysis, we showed that the plasmid-based ts-mutant ΔfabA/pTS-fabA failed to grow at a restrictive temperature, consistent with the observation by Hoang and Schweizer (T. T. Hoang, H. P. Schweizer, J Bacteriol 179:5326–5332, 1997, https://doi.org/10.1128/jb.179.17.5326-5332.1997), and expanded on this by showing that ΔfabA exhibited curved cell morphology. On the other hand, strong induction of fabA-OE or PA3645|fabZ-OE impeded the growth of cells displaying oval morphology. Suppressor analysis revealed a mutant sup gene that suppressed a growth defect but not cell morphology of ΔfabA. Genome resequencing and transcriptomic profiling of sup identified PA0286|desA, whose promoter carried a single-nucleotide polymorphism (SNP), and transcription was significantly upregulated (level increase of >2-fold, P < 0.05). By integration of the SNP-bearing promoter-controlled desA gene into the chromosome of ΔfabA/pTS-fabA, we showed that the SNP is sufficient for ΔfabA to phenocopy the sup mutant. Furthermore, mild induction of the araC-PBAD-controlled desA gene but not desB rescued ΔfabA. These results validated that mild overexpression of desA fully suppressed the lethality but not the curved cell morphology of ΔfabA. Similarly, Zhu et al. (Zhu K, Choi K-H, Schweizer HP, Rock CO, Zhang Y-M, Mol Microbiol 60:260–273, 2006, https://doi.org/10.1111/j.1365-2958.2006.05088.x) showed that multicopy desA partially alleviated the slow growth phenotype of ΔfabA, the difference in which was that ΔfabA was viable. Taken together, our results demonstrate that fabA is essential for aerobic growth. We propose that the plasmid-based ts-allele is useful for exploring the genetic suppression interaction of essential genes of interest in P. aeruginosa. IMPORTANCE Pseudomonas aeruginosa is an opportunistic pathogen whose multidrug resistance demands new drug development. Fatty acids are essential for viability, and essential genes are ideal drug targets. However, the growth defect of essential gene mutants can be suppressed. Suppressors tend to be accumulated during the construction of essential gene deletion mutants, hampering the genetic analysis. To circumvent this issue, we constructed a deletion allele of fabA in the presence of a native promoter-controlled complementary copy in the ts-plasmid. In this analysis, we showed that ΔfabA/pTS-fabA failed to grow at a restrictive temperature, supporting its essentiality. Suppressor analysis revealed desA, whose promoter carried a SNP and whose transcription was upregulated. We validated that both the SNP-bearing promoter-controlled and regulable PBAD promoter-controlled desA suppressed the lethality of ΔfabA. Together, our results demonstrate that fabA is essential for aerobic growth. We propose that plasmid-based ts-alleles are suitable for genetic analysis of essential genes of interest.

Emotional competence (EC) has been documented as one of the most influential resource to protect parents from parental burnout (PB). However, the dimensions of EC have inconsistent effects across studies: while intrapersonal EC consistently had a protective effect on PB, interpersonal EC was paradoxically documented both as a risk factor and a protective factor. Relying on third-variable effect analyses on two independent datasets (842 Belgian parents and 377 Polish parents), this pre-registered study found that (1) intrapersonal EC drives the protective effect of interpersonal EC, and (2) after being adjusted for intrapersonal EC, interpersonal EC does not have significant effect on PB in the Polish sample and even becomes detrimental in the Belgian sample. These findings highlight the importance of examining the unique effect of each EC dimension while controlling for the others. Besides, it also implicates interventions aimed at improving EC to reduce PB should focus on intrapersonal EC. [ABSTRACT FROM AUTHOR]

Anthocyanin pigments furnish a powerful visual output of the stress and metabolic status of Arabidopsis thaliana plants. Essential for pigment accumulation is TRANSPARENT TESTA19 (TT19), a glutathione S-transferase proposed to bind and stabilize anthocyanins, participating in their vacuolar sequestration, a function conserved across the flowering plants. Here, we report the identification of genetic suppressors that result in anthocyanin accumulation in the absence of TT19. We show that mutations in RDR6, SGS3, or DCL4 suppress the anthocyanin defect of tt19 by pushing carbon towards flavonoid biosynthesis. This effect is not unique to tt19 and extends to at least one other anthocyanin pathway gene mutant. This synergy between mutations in components of the RDR6-SGS3-DCL4 siRNA system and the flavonoid pathway reveals genetic/epigenetic mechanisms regulating metabolic fluxes.

Mediator 12 (MED12) and MED13 are components of the Mediator multi-protein complex, that facilitates the initial steps of gene transcription. Here, in an Arabidopsis mutant screen, we identify MED12 and MED13 as positive gene regulators, both of which contribute broadly to morc1 de-repressed gene expression. Both MED12 and MED13 are preferentially required for the expression of genes depleted in active chromatin marks, a chromatin signature shared with morc1 re-activated loci. We further discover that MED12 tends to interact with genes that are responsive to environmental stimuli, including light and radiation. We demonstrate that light-induced transient gene expression depends on MED12, and is accompanied by a concomitant increase in MED12 enrichment during induction. In contrast, the steady-state expression level of these genes show little dependence on MED12, suggesting that MED12 is primarily required to aid the expression of genes in transition from less-active to more active states.

The SCARECROW (SCR) transcription factor plays a key role in plant growth and development. However, we know very little about the role of SCR regulated pathways in plant development. Here, we used the homozygous scr1 mutant Arabidopsis thaliana (Wassilewskija ecotype), which had a T-DNA insertion in the SCR coding region and lacks a detectable SCR transcript. This scr1 mutant has a determinate mode of root growth, shoot agravitropism and abnormal internal architecture in all organs examined. To screen for mutants that suppress the scr1 abnormal phenotypes, we exposed homozygous scr1 seeds to ethyl methane sulphonate (EMS) mutagen. Upon growth out of these mutagenized seeds, thirteen suppressor mutant-harboring strains were identified. All thirteen suppressor-harboring strains were homozygous for scr1 and lacked the SCR transcript. Ten scr hypocotyl gravitropic suppressor lines showed improved hypocotyl gravitropic response. These ten suppressors fall into six complementation groups suggesting six different gene loci. Similarly, three independent scr root length suppressor lines rescued only the root growth phenotype and fell into three complementation groups, suggesting the involvement of three different gene loci. These suppressors might identify novel functions of the SCR gene in plant development.

The function of Vasohibin-1 (VASH1) in human cancer has not been thoroughly or comprehensively examined. Here, we identified the tumor suppressor part of VASH1 across cancers, including epithelial ovarian tumors. Our study carefully contrasted the expression of VASH1 in pancancer and nontumorous tissues in a public database to explore its regulatory role in clinical prognosis, diagnosis, tumor purity, and immune cell infiltration. Next, we explored the antitumor mechanism of VASH1 through drug sensitivity, functional enrichment, and phenotypic experiments in ovarian cancer. Research suggests that the expression of VASH1 in neoplastic tissues is lower than that in normal tissues. VASH1 affects the OS and RFS of several tumor types. In addition, VASH1 expression resulted in a high OS and RFS in the diagnosis of tumor and nontumor tissues and negatively regulated tumor purity. Moreover, VASH1 controls the tumor microenvironment by regulating immunocyte infiltration. In ovarian cancer, VASH1 can serve as a biomarker to estimate the efficacy of chemotherapy. Functional enrichment analysis suggests that VASH1 plays a tumor suppressor role by regulating the extracellular matrix receptor pathway. VASH1 inhibition of the malignant phenotype of ovarian cancer cells was further confirmed by in vivo experiments. These results indicate that VASH1 acts as a cancer-inhibiting factor and potential therapeutic target in ovarian cancer. [ABSTRACT FROM AUTHOR]

In this study, we identified a new citrus vein enation virus (CVEV) isolate (named CVEV-DT1) through sRNA high-throughput sequencing and traditional sequencing. Phylogenetic analysis based on whole genome sequences of all known CVEV isolates revealed that CVEV-DT1 was in an evolutionary branch with other isolates from China. Molecular variation analysis showed that the single nucleotide variability along CVEV full-length sequences was less than 8%, with more transitions (60.55%) than transversions (39.43%), indicating a genetically homogeneous CVEV population. In addition, non-synonymous nucleotide mutations mainly occurred in ORF1 and ORF2. Based on disorder analysis of all encoded ORF by CVEV-DT1, we identified that the CVEV-DT1 coat protein (CP) formed spherical granules, mainly in the cell nucleus and partly throughout the cytoplasm, with liquid properties through subcellular localization and photobleaching assay. Furthermore, we also confirmed that the CVEV P0 protein has weak post-transcriptional RNA-silencing suppressor activity and could elicit a strong hypersensitive response (HR) in tobacco plants. Collectively, to the best of our knowledge, our study was the first to profile the genomic variation in all the reported CVEV isolates and reveal the functions of CVEV-DT1-encoded proteins. [ABSTRACT FROM AUTHOR]

The toxic components and odorous components in the fumes of petroleum asphalt are introduced in detail, and some suggestions for the establishment of the standard and specification of the toxic components and odorous components in the fumes of petroleum asphalt are put forward・ The toxic components in petroleum asphalt smoke are mainly 2 〜6 rings of polycyclic aromatic hydrocarbons (PAHs), including benzo [a] anthracene, benzo [a] pyrene and benzo [e] pyrene, which have been shown to be carcinogenic to human body and toxic to nervous system・ The odor group of petroleum asphalt flue gas is divided into sulfur and nitrogen compounds, including thiophene similar substance and sulfoxide, and also contains a small amount of indole nitrogen odor pollutants・ The research status of adsorption type fumes suppressor, oxidation type fumes suppressor and polymer fumes suppressor in petroleum asphalt suppression technology is introduced and evaluated・ In add让ion, the research expectation of petroleum asphalt fumes suppressor is predicted. [ABSTRACT FROM AUTHOR]

The SCARECROW (SCR) transcription factor plays a key role in plant growth and development. However, we know very little about the role of SCR regulated pathways in plant development. Here, we used the homozygous scr1 mutant Arabidopsis thaliana (Wassilewskija ecotype), which had a T-DNA insertion in the SCR coding region and lacks a detectable SCR transcript. This scr1 mutant has a determinate mode of root growth, shoot agravitropism and abnormal internal architecture in all organs examined. To screen for mutants that suppress the scr1 abnormal phenotypes, we exposed homozygous scr1 seeds to ethyl methane sulphonate (EMS) mutagen. Upon growth out of these mutagenized seeds, thirteen suppressor mutant-harboring strains were identified. All thirteen suppressor-harboring strains were homozygous for scr1 and lacked the SCR transcript. Ten scr hypocotyl gravitropic suppressor lines showed improved hypocotyl gravitropic response. These ten suppressors fall into six complementation groups suggesting six different gene loci. Similarly, three independent scr root length suppressor lines rescued only the root growth phenotype and fell into three complementation groups, suggesting the involvement of three different gene loci. These suppressors might identify novel functions of the SCR gene in plant development. [ABSTRACT FROM AUTHOR]

Simple Summary: Abnormal expression of miRNA is often observed in cancer. MiR-101 is closely associated with tumor initiation and progression. In this review, we summarized new findings regarding the role of miR-101 in cancer and the potential mechanisms of targeted gene degradation and microenvironmental regulation. MiRNAs are small single-stranded non-coding RNAs. MiRNA contributes to the transcriptional and post-transcriptional regulation of mRNA in different cell types, including mRNA transcription inhibition and mRNA decay and phenotypes via the effect of several essential oncogenic processes and tumor microenvironment. MiR-101 is a highly conserved miRNA that was found to alter the expression in various human cancers. MiR-101 has been reported to have tumor oncogenic and suppressive effects to regulate tumorigenesis and tumor progression. In this review, we summarize the new findings about the roles of miR-101 in cancers and the underlying mechanisms of targeting genes degradation and microenvironment regulation, which will improve biological understanding and design of novel therapeutics. [ABSTRACT FROM AUTHOR]

Gross Chromosomal Rearrangements (GCRs) play an important role in human diseases, including cancer. Although most of the nonessential Genome Instability Suppressing (GIS) genes in Saccharomyces cerevisiae are known, the essential genes in which mutations can cause increased GCR rates are not well understood. Here 2 S. cerevisiae GCR assays were used to screen a targeted collection of temperature-sensitive mutants to identify mutations that caused increased GCR rates. This identified 94 essential GIS (eGIS) genes in which mutations cause increased GCR rates and 38 candidate eGIS genes that encode eGIS1 protein-interacting or family member proteins. Analysis of TCGA data using the human genes predicted to encode the proteins and protein complexes implicated by the S. cerevisiae eGIS genes revealed a significant enrichment of mutations affecting predicted human eGIS genes in 10 of the 16 cancers analyzed.

Background & aimsMutations at hotspots in GNAS, which encodes stimulatory G-protein, α subunits, are detected in approximately 60% of intraductal papillary mucinous neoplasms (IPMNs) of the pancreas. We generated mice with KRAS-induced IPMNs that also express a constitutively active form of GNAS in pancreas and studied tumor development.MethodsWe generated p48-Cre; LSL-KrasG12D; Rosa26R-LSL-rtTA-TetO-GnasR201C mice (Kras;Gnas mice); pancreatic tissues of these mice express activated KRAS and also express a mutant form of GNAS (GNASR201C) upon doxycycline administration. Mice that were not given doxycycline were used as controls, and survival times were compared by Kaplan-Meier analysis. Pancreata were collected at different time points after doxycycline administration and analyzed by histology. Pancreatic ductal adenocarcinomas (PDACs) were isolated from mice and used to generate cell lines, which were analyzed by reverse transcription polymerase chain reaction, immunoblotting, immunohistochemistry, and colony formation and invasion assays. Full-length and mutant forms of yes-associated protein (YAP) were expressed in PDAC cells. IPMN specimens were obtained from 13 patients with IPMN undergoing surgery and analyzed by immunohistochemistry.ResultsAll Kras;Gnas mice developed pancreatic cystic lesions that resemble human IPMNs; the grade of epithelial dysplasia increased with time. None of the control mice developed cystic lesions. Approximately one third of Kras;Gnas mice developed PDACs at a median of 30 weeks after doxycycline administration, whereas 33% of control mice developed PDACs. Expression of GNASR201C did not accelerate the development of PDACs compared with control mice. However, the neoplasms observed in Kras;Gnas mice were more differentiated, and expressed more genes associated with ductal phenotypes, than in control mice. PDACs isolated from Kras;Gnas mice had activation of the Hippo pathway; in cells from these tumors, phosphorylated YAP1 was sequestered in the cytoplasm, and this was also observed in human IPMNs with GNAS mutations. Sequestration of YAP1 was not observed in PDAC cells from control mice.ConclusionsIn mice that express activated KRAS in the pancreas, we found expression of GNASR201C to cause development of more differentiated tumors, with gene expression pattern associated with the ductal phenotype. Expression of mutant GNAS caused phosphorylated YAP1 to be sequestered in the cytoplasm, altering tumor progression.

Simple Summary: Gastric cancer is a common upper gastrointestinal tumor. Death-associated protein kinase (DAPK1) was found to participate in the development of various malignant tumors. However, there are few reports on DAPK1 in gastric cancer. In this study, we found that the expression of DAPK1 was up-regulated in gastric cancer tissues. Survival analysis showed that low expression of DAPK1 was a favorable prognostic factor of overall survival and disease-free survival for gastric cancer patients. The functional experiments demonstrated that DAPK1 can promote the migration and invasion of gastric cancer cells. Meanwhile, a risk score module for four DAPK1-related genes was constructed and validated. In conclusion, these results indicated that DAPK1 can promote gastric cancer cell migration and invasion and established a signature of four DAPK1-related genes for gastric cancer that could independently predict the survival of gastric cancer patients. Gastric cancer (GC) is a common upper gastrointestinal tumor. Death-associated protein kinase (DAPK1) was found to participate in the development of various malignant tumors. However, there are few reports on DAPK1 in gastric cancer. In this study, the TCGA and GEO datasets were used to explore the expression and role of DAPK1 in gastric cancer. The functions of DAPK1 in gastric cancer were determined by proliferation, migration and invasion assays. In addition, genes co-expressed with DAPK1 in gastric cancer were estimated through the WGCNA and correlation analysis. A DAPK1-related gene prognostic model was constructed using the Cox regression and lasso analyses. The expression of DAPK1 was significantly up-regulated in gastric cancer tissues. Kaplan–Meier analysis showed that low expression of DAPK1 was a favorable prognostic factor of overall survival and disease-free survival for gastric cancer patients. Functional experiments demonstrated that DAPK1 can promote the migration and invasion of gastric cancer cells. WGCNA, correlation analysis, Cox regression, and lasso analyses were applied to construct the DAPK1-related prognostic model. The prognostic value of this prognostic model of DAPK1-related genes was further successfully validated in an independent database. Our results indicated that DAPK1 can promote gastric cancer cell migration and invasion and established four DAPK1-related signature genes for gastric cancer that could independently predict the survival of GC patients. [ABSTRACT FROM AUTHOR]

Metabolic dysregulation leading to sugar-phosphate accumulation is toxic in organisms ranging from bacteria to humans. By comparing two models of sugar-phosphate toxicity in Saccharomyces cerevisiae, we demonstrate that toxicity occurs, at least in part, through multiple, isomer-specific mechanisms, rather than a single general mechanism.

Light utilization is finely tuned in photosynthetic organisms to prevent cellular damage. The dissipation of excess absorbed light energy, a process termed nonphotochemical quenching (NPQ), plays an important role in photoprotection. Little is known about the sustained or slowly reversible form(s) of NPQ and whether they are photoprotective, in part due to the lack of mutants. The Arabidopsis thaliana suppressor of quenching1 (soq1) mutant exhibits enhanced sustained NPQ, which we term qH. To identify molecular players involved in qH, we screened for suppressors of soq1 and isolated mutants affecting either chlorophyllide a oxygenase or the chloroplastic lipocalin, now renamed plastid lipocalin (LCNP). Analysis of the mutants confirmed that qH is localized to the peripheral antenna (LHCII) of photosystem II and demonstrated that LCNP is required for qH, either directly (by forming NPQ sites) or indirectly (by modifying the LHCII membrane environment). qH operates under stress conditions such as cold and high light and is photoprotective, as it reduces lipid peroxidation levels. We propose that, under stress conditions, LCNP protects the thylakoid membrane by enabling sustained NPQ in LHCII, thereby preventing singlet oxygen stress.

Firespike leafroll-associated virus (FLRaV) is a major pathogen associated with firespike (Odontonema tubaeforme) leafroll disease. Phylogenetic analysis showed that FLRaV possesses typical traits of subgroup II members of ampeloviruses, but encodes two additional proteins, P25 and P37. Here, we determined the microfilament localization of P25 protein. Posttranscriptional gene silencing (PTGS) assay showed that both FLRaV P25 and P37 were able to suppress the local and systemic PTGS and FLRaV P25 was capable of suppressing the green fluorescent protein (GFP) gene silencing triggered by both sense RNA-induced PTGS (S-PTGS) and inverted repeat RNA-induced PTGS (IR-PTGS). In contrast, FLRaV P37 was only able to inhibit the GFP silencing triggered by the S-PTGS but not the IR-PTGS. In the transcriptional gene silencing (TGS) assay, only FLRaV P25 was found to be able to reverse established TGS-mediated silencing of GFP in 16-TGS plants. We also found that FLRaV P25 could aggravate the disease symptom and viral titer of potato virus X in N. benthamiana. These results suggest that FLRaV P25 and P37 may have crucial roles in overcoming host RNA silencing, which provides key insights into our understanding of the molecular mechanisms underlying FLRaV infection. [ABSTRACT FROM AUTHOR]

Over 300 essential genes are predicted using transposon sequencing in the genome of Pseudomonas aeruginosa. However, methods for reverse genetic analysis of essential genes are scarce. To address this issue, we developed a three-step protocol consisting of integration of deletion plasmid, introduction of temperature-sensitive rescue plasmid, and excision of integrated-deletion plasmid to construct the plasmid-based temperature-sensitive allele of essential genes. Using PA0006 as an example, we showed that PA0006(Ts) exhibited wild-type cell morphology at permissive temperature but filamentous form at restrictive temperatures. We further showed that the glycerol-mannoheptose-bisphosphate phosphatase GmhB in Escherichia coli shared 32.4% identity with that of PA0006p and functionally complemented the defect of PA0006(Ts) at 42°C. SDS-PAGE and Western blotting indicated the presence and absence of the complete core lipopolysaccharide (LPS) and B-band O-antigen in PA0006(Ts) at 30 and 42°C, respectively. An isolated suppressor sup displayed wild-type-like cell morphology but no complete core LPS or O-antigen. Genome rese-quencing together with comparative transcriptomic profiling identified a candidate suppressor fructose-bisphosphate phosphatase in which the promoter harbored a SNP and the transcription level was not downregulated at 42°C compared to 30°C in sup. It was further validated that fbp overexpression suppressed the lethality of PA0006(Ts) at 42°C. Taken together, our results demonstrate that PA0006 plays a role in regulation of cell morphology and biosynthesis of core LPS. This three-step protocol for construction of conditional lethal allele in P. aeruginosa should be widely applicable for genetic analysis of other essential genes of interest, including analysis of bypass suppressibility. IMPORTANCE Microbial essential genes encode nondispensable function for cell growth and therefore are ideal targets for the development of new drugs. Essential genes are readily identified using transposon-sequencing technology at the genome scale. However, genetic analysis of essential genes of interest was hampered by limited methodologies. To address this issue, we developed a three-step protocol for construction of conditional allele of essential genes in the opportunistic pathogen Pseudomonas aeruginosa. Using PA0006 as an example, we demonstrated that the plasmid-based PA0006(Ts) mutant exhibited defects in regulation of cell morphology, formation of intact core LPS, and attachment of the O-antigen at restrictive temperatures but not at permissive temperatures. A suppressor of PA0006(Ts) was isolated through spontaneous mutations and showed restored cell morphology but not core oligosaccharide or O-antigen. This method should be widely applicable for pheno-type and suppressibility analyses of other essential genes of interest in P. aeruginosa. [ABSTRACT FROM AUTHOR]

Integrin plays a crucial role in the attachment of cells to the extracellular matrix. Integrin recruits many proteins intracellularly, including a 4-protein complex (kindlin, ILK, PINCH, and parvin). Caenorhabditis elegans muscle provides an excellent model to study integrin adhesion complexes. In Caenorhabditis elegans, UNC-112 (kindlin) binds to the cytoplasmic tail of PAT-3 (b-integrin) and to PAT-4 (ILK). We previously reported that PAT-4 binding to UNC-112 is essential for the binding of UNC-112 to PAT-3. Although there are crystal structures for ILK and a kindlin, there is no co-crystal structure available. To understand the molecular interaction between PAT-4 and UNC-112, we took a genetic approach. First, using a yeast 2-hybrid method, we isolated mutant PAT-4 proteins that cannot bind to UNC-112 and then isolated suppressor mutant UNC-112 proteins that restore interaction with mutant PAT-4 proteins. Second, we demonstrated that these mutant PAT-4 proteins cannot localize to attachment structures in nematode muscle, but upon co-expression of an UNC-112 suppressor mutant protein, mutant PAT- 4 proteins could localize to attachment structures. Third, overexpression of a PAT-4 mutant results in the disorganization of adhesion plaques at muscle cell boundaries and co-expression of the UNC-112 suppressor mutant protein alleviates this defect. Thus, we demonstrate that UNC-112 binding to PAT-4 is required for the localization and function of PAT-4 in integrin adhesion complexes in vivo. The missense mutations were mapped onto homology models of PAT-4 and UNC-112, and taking into account previously isolated mutations, we suggest a surface of PAT-4 that binds to UNC-112. [ABSTRACT FROM AUTHOR]

SUMMARY: In rice (Oryza sativa), the lemma and palea protect the internal organs of the floret，provide nutrients for seed development, and determine grain size. We previously revealed that a trans‐acting small interfering RNA targeting AUXIN RESPONSE FACTORS (tasiR‐ARF) regulates lemma polarity establishment via post‐transcriptional repression of AUXIN RESPONSE FACTORS (ARFs) in rice. TasiR‐ARF formation requires RNA‐DEPENDENT RNA POLYMERASE 6 (RDR6). However, the underlying molecular mechanism of the tasiR‐ARF–ARF regulon in lemma development remains unclear. Here, by genetic screening for suppressors of the thermosensitive mutant osrdr6‐1, we identified three suppressors, huifu 1 (hf1), hf9, and hf17. Mapping‐by‐sequencing revealed that HF1 encodes a MYB transcription factor belonging to the KANADI1 family. The hf1 mutation partially rescued the osrdr6‐1 lemma defect but not the defect in tasiR‐ARF levels. DNA affinity purification sequencing analysis identified 17 725 OsKANADI1‐associated sites, most of which contain the SPBP‐box binding motif (RGAATAWW) and are located in the promoter, protein‐coding, intron, and intergenic regions. Moreover, we found that OsKANADI1 could directly bind to the intron of OsARF3a in vitro and in vivo and promote OsARF3a expression at the transcriptional level. In addition, hf9 and hf17 are intragenic suppressors containing mutations in OsRDR6 that partially rescue tasiR‐ARF levels by restoring OsRDR6 protein levels. Collectively, our results demonstrate that OsKANADI1 and tasiR‐ARFs synergistically maintain the proper expression of OsARF3a and thus contribute to rice lemma development. Significance Statement: The lemma and palea are unique and important organs for rice, providing nutrients and protection for floret and seed development, and directly affecting grain size. Our findings show that the expression of OsARF3a was controlled by both OsKANADI1 and via RDR6 at the transcription and post‐transcription levels, thus ensuring the normal development of glumes in rice. [ABSTRACT FROM AUTHOR]

Sensory organ glia surround neuronal receptive endings (NREs), forming a specialized compartment important for neuronal activity, and reminiscent of glia-ensheathed synapses in the central nervous system. We previously showed that DAF-6, a Patched-related protein, is required in glia of the C. elegans amphid sensory organ to restrict sensory compartment size. LIT-1, a Nemo-like kinase, and SNX-1, a retromer component, antagonize DAF-6 and promote compartment expansion. To further explore the machinery underlying compartment size control, we sought genes whose inactivation restores normal compartment size to daf-6 mutants. We found that mutations in igdb-2, encoding a single-pass transmembrane protein containing Ig-like and fibronectin type III domains, suppress daf-6 mutant defects. IGDB-2 acts in glia, where it localizes to glial membranes surrounding NREs, and, together with LIT-1 and SNX-1, regulates compartment morphogenesis. Immunoprecipitation followed by mass spectrometry demonstrates that IGDB-2 binds to LGC-34, a predicted ligand-gated ion channel, and lgc-34 mutations inhibit igdb-2 suppression of daf-6. Our findings reveal a novel membrane protein complex and suggest possible mechanisms for how sensory compartment size is controlled.

MicroRNAs (miRNAs) are a group of non‑coding RNAs that exert master regulatory functions in post‑-transcriptional gene expression. Accumulating evidence shows that miRNAs can either promote or suppress tumorigenesis by regulating different target genes or pathways and may be involved in the occurrence of carcinoma. miR‑409‑3p is dysregulated in a variety of malignant cancers. It plays a fundamental role in numerous cellular biological processes, such as cell proliferation, apoptosis, migration, invasion, autophagy, angiogenesis and glycolysis. In addition, studies have shown that miR‑409‑3p is expected to become a non‑invasive biomarker. Identifying the molecular mechanisms underlying miR‑409‑3p‑mediated tumor progression will help investigate miR‑409‑3p‑based targeted therapy for human cancers. The present review comprehensively summarized the recently published literature on miR‑409‑3p, with a focus on the regulation and function of miR‑409‑3p in various types of cancer, and discussed the clinical implications of miR‑409‑3p, providing new insight for the diagnosis and treatment of cancers.

The metastatic suppressor BRMS1 interacts with critical steps of the metastatic cascade in many cancer entities. As gliomas rarely metastasize, BRMS1 has mainly been neglected in glioma research. However, its interaction partners, such as NFκB, VEGF, or MMPs, are old acquaintances in neurooncology. The steps regulated by BRMS1, such as invasion, migration, and apoptosis, are commonly dysregulated in gliomas. Therefore, BRMS1 shows potential as a regulator of glioma behavior. By bioinformatic analysis, in addition to our cohort of 118 specimens, we determined BRMS1 mRNA and protein expression as well as its correlation with the clinical course in astrocytomas IDH mutant, CNS WHO grade 2/3, and glioblastoma IDH wild-type, CNS WHO grade 4. Interestingly, we found BRMS1 protein expression to be significantly decreased in the aforementioned gliomas, while BRMS1 mRNA appeared to be overexpressed throughout. This dysregulation was independent of patients’ characteristics or survival. The protein and mRNA expression differences cannot be finally explained at this stage. However, they suggest a post-transcriptional dysregulation that has been previously described in other cancer entities. Our analyses present the first data on BRMS1 expression in gliomas that can provide a starting point for further investigations.

Many plant viruses express suppressor proteins (VSRs) that can inhibit RNA silencing, a central component of antiviral plant immunity. The most common activity of VSRs is the high-affinity binding of virus-derived siRNAs and thus their sequestration from the silencing process. Since siRNAs share large homologies with miRNAs, VSRs like the Tombusvirus p19 may also bind miRNAs and in this way modulate cellular gene expression at the post-transcriptional level. Interestingly, the binding affinity of p19 varies considerably between different miRNAs, and the molecular determinants affecting this property have not yet been adequately characterized. Addressing this, we analyzed the binding of p19 to the miRNAs 162 and 168, which regulate the expression of the important RNA silencing constituents Dicer-like 1 (DCL1) and Argonaute 1 (AGO1), respectively. p19 binds miRNA162 with similar high affinity as siRNA, whereas the affinity for miRNA168 is significantly lower. We show that specific molecular features, such as mismatches and 'G–U wobbles' on the RNA side and defined amino acid residues on the VSR side, mediate this property. Our observations highlight the remarkable adaptation of VSR binding affinities to achieve differential effects on host miRNA activities. Moreover, they show that even minimal changes, i.e., a single base pair in a miRNA duplex, can have significant effects on the efficiency of the plant antiviral immune response. [ABSTRACT FROM AUTHOR]

Communication between interacting organisms via bioactive molecules is widespread in nature and plays key roles in diverse biological processes. Small RNAs (sRNAs) can travel between host plants and filamentous pathogens to trigger transkingdom RNA interference (RNAi) in recipient cells and modulate plant defense and pathogen virulence. However, how fungal pathogens counteract transkingdom antifungal RNAi has rarely been reported. Here we show that a secretory protein VdSSR1 (secretory silencing repressor 1) from Verticillium dahliae, a soil-borne phytopathogenic fungus that causes wilt diseases in a wide range of plant hosts, is required for fungal virulence in plants. VdSSR1 can translocate to plant nucleus and serve as a general suppressor of sRNA nucleocytoplasmic shuttling. We further reveal that VdSSR1 sequesters ALY family proteins, adaptors of the TREX complex, to interfere with nuclear export of the AGO1–microRNA (AGO1–miRNA) complex, leading to a great attenuation in cytoplasmic AGO1 protein and sRNA levels. With this mechanism, V. dahliae can suppress the accumulation of mobile plant miRNAs in fungal cells and succedent transkingdom silencing of virulence genes, thereby increasing its virulence in plants. Our findings reveal a mechanism by which phytopathogenic fungi antagonize antifungal RNAidependent plant immunity and expand the understanding on the complex interaction between host and filamentous pathogens. [ABSTRACT FROM AUTHOR]

While most missense suppressors have very narrow specificities and only suppress the allele against which they were isolated, the sumA missense suppressor from Salmonella enterica serovar Typhimurium is a promiscuous or broad-acting missense suppressor that suppresses numerous missense mutants. The sumA missense suppressor was identified as a glyV tRNA Gly3(GAU/C) missense suppressor that can recognize GAU or GAC aspartic acid codons and insert a glycine amino acid instead of aspartic acid. In addition to rescuing missense mutants caused by glycine to aspartic acid changes as expected, sumA could also rescue a number of other missense mutants as well by changing a neighboring (contacting) aspartic acid to glycine, which compensated for the other amino acid change. Thus the ability of sumA to rescue numerous missense mutants was due in part to the large number of glycine codons in genes that can be mutated to an aspartic acid codon and in part to the general tolerability and/or preference for glycine amino acids in proteins. Because the glyV tRNA Gly3(GAU/C) missense suppressor has also been extensively characterized in Escherichia coli as the mutA mutator, we demonstrated that all gain-of-function mutants isolated in a glyV tRNA Gly3(GAU/C) missense suppressor are transferable to a wild-type background and thus the increased mutation rates, which occur in glyV tRNA Gly3(GAU/C) missense suppressors, are not due to the suppression of these mutants.

Jiali Wang,1,2 Tiangong Wang,2 Shiyun Hu,2 Jinyun Li,1 Chao Ni,1 Meng Ye1 1Department of Oncology and Hematology, The Affiliated Hospital of Medical School, Ningbo University, Ningbo, People’s Republic of China; 2School of Medicine, Ningbo University, Ningbo, People’s Republic of ChinaCorrespondence: Meng YeDepartment of Oncology and Hematology, The Affiliated Hospital of Medical School, Ningbo University, 247 Renming Road, Ningbo, Zhejiang, 315000, People’s Republic of ChinaTel +86 19857840350Email yemeng@nbu.edu.cnPurpose: Circular RNAs are novel endogenous RNAs, which are considered to play a role in tumorigenesis. Nevertheless, the role as well as clinical diagnostic value of most circular RNAs in colorectal cancer are still unclear.Materials and Methods: We investigated the circular RNA microarray containing expression profiles in samples of colorectal cancer patients by bioinformatics. The consequence indicated that hsa_circ_0043278 was strongly downregulated. We then measured the expression level of hsa_circ_0043278 in tissue samples of colorectal cancer by quantitative real-time polymerase chain reaction. Besides, we also explored the expression condition of the circular RNA in colorectal cancer cell lines including HCT116, SW620, and SW480. Cell counting kit-8, colony formation, and transwell assays, as well as flow cytometry, were applied to detect changes in cell proliferation, migration, apoptosis, and cell cycle progression.Results: We discovered that circular RNA hsa_circ_0043278 was significantly downregulated in tumor samples (P < 0.0001) as well as cell lines (P < 0.05). The value of the area under the receiver operating characteristic curve was 0.71, with a sensitivity of 0.72 and specificity of 0.70 (P = 0.0006). Moreover, we found that overexpression of hsa_circ_0043278 suppressed proliferation and migratory abilities while promoting apoptosis in colorectal cancer cells.Conclusion: Our findings revealed that hsa_circ_0043278 inhibited the tumorigenesis of colorectal cancer and could be a potential biomarker for colorectal cancer diagnosis. Besides, it hopes to become a target for treatment.Keywords: circular RNAs, apoptosis, suppressor, molecular marker, tumorigenesis

This study explores the structure-activity relationship of copper electroplating suppressors, with a focus on alkyl-terminated polyethylene glycols (PEGs), to better understand their efficacy in electronic interconnect fabrication. Electrochemical analysis demonstrated that an increase in the molecular weight of PEG suppressors significantly raises the electrochemical polarization overpotential during copper electrodeposition. Additionally, alkyl terminal capping was observed to amplify this polarization effect, with the extent of overpotential increase directly correlating with the alkyl chain length. However, the advantageous impact of molecular weight on overpotential diminishes at higher molecular weights. In contrast to the electrochemical results, through-hole electroplating experiments revealed that a suppressor with a molecular weight of 1 k and butyl termination exhibits superior through-hole throwing power. Further analysis indicated that a greater proportion of alkyl functional groups within the suppressor's molecular chain enhances uniform electroplating capabilities. The study also suggests that measuring contact angles, as an expression of hydrophobicity, could provide a more efficient and precise method for evaluating suppressor performance. [Display omitted] • Alkyl-terminated PEG suppressors exhibit higher polarization and throwing power in acid copper electrodeposition. • Alkyls increase PEGs' hydrophobicity, crucial for uniform through-hole metallization. • Contact angle measurements efficiently and accurately evaluate suppressors, surpassing traditional techniques. [ABSTRACT FROM AUTHOR]