The histone methyltransferase Set7/9 promotes myoblast differentiation and myofibril assembly

Set7 associates with the MyoD transcription factor to enhance expression of genes required for muscle differentiation

Functional Analysis of General Odorant Binding Protein 2 from the Meadow Moth, Loxostege sticticalis L. (Lepidoptera: Pyralidae)

Odorant binding proteins play a crucial role in transporting semiochemicals across the sensillum lymph to olfactory receptors within the insect antennal sensilla. In this study, the general odorant binding protein 2 gene was cloned from the antennae of Loxostege sticticalis, using reverse transcription PCR and rapid amplification of cDNA ends. Recombinant LstiGOBP2 was expressed in Escherichia coli and purified by Ni ion affinity chromatography. Real-time PCR assays indicated that LstiGOBP2 mRNA is expressed mainly in adult antennae, with expression levels differing with developmental age. Ligand-binding experiments using N-phenyl-naphthylamine (1-NPN) as a fluorescent probe demonstrated that the LstiGOBP2 protein has binding affinity to a broad range of odorants. Most importantly, trans-11-tetradecen-1-yl acetate, the pheromone component of Loxostege sticticalis, and trans-2-hexenal and cis-3-hexen-1-ol, the most abundant plant volatiles in essential oils extracted from host plants, had high binding affinities to LstiGOBP2 and elicited strong electrophysiological responses from the antennae of adults

Identification and Molecular Characterization of a Chitin-Binding Protein from the Beet Webworm, Loxostege sticticalis L.

As the first crucial barrier in the midgut of insects, the peritrophic membrane (PM) plays an important role in preventing external invasion. PM proteins, as the major components of the PM, determine the structure and function of this membrane. A new PM protein, named LstiCBP, from the PM of Loxostege sticticalis larvae was identified using cDNA library screening. The full cDNA of LstiCBP is 2606 bp in length and contains a 2403 bp ORF that encodes an 808-amino acid preprotein with a 15-amino acid as signal peptide. The deduced protein sequence of the cDNA contains 8 cysteine-rich chitin-binding domains (CBDs). Recombinant LstiCBP was successfully expressed in BL21 cells using recombinant plasmid DNA and showed high chitin-binding activity. LstiCBP expression was detected in the midgut at both the transcriptional and translational levels; however, the biochemical and physiological functions of LstiCBP in L. sticticalis require further investigation

Potential cooperations between odorant-binding proteins of the scarab beetle Holotrichia oblita Faldermann (Coleoptera: Scarabaeidae).

It was previously thought that the odorant binding proteins (OBPs) in the sensillum lymph might serve as carriers, which could carry lipophilic odorant molecules to olfactory receptors. In this study, two novel OBP genes of the scarab beetle Holotrichia oblita were screened using an antennal cDNA library. The full cDNA of HoblOBP3 and HoblOBP4 was cloned using reverse transcription PCR and rapid amplification of the cDNA ends. Homology modeling of both OBPs was performed using SWISS-MODEL on-line tools. Next, the two OBPs were expressed in Escherichia coli and purified using Ni ion affinity chromatography. The ligand-binding properties of HoblOBP3 and HoblOBP4 in 42 ligands respectively were measured using the fluorescence probe N-phenyl-naphthylamine (1-NPN). The results obtained from competitive binding assays demonstrated that HoblOBP4 showed a broader range of binding affinities to the test compounds, while HoblOBP3 displays more specific binding affinity. Furthermore, other OBPs and CSPs were expressed in Escherichia coli and purified using Ni ion affinity chromatography. Binding curves were measured for binary mixtures of OBPs and CSPs using 1-NPN, and the Scatchard plots exhibited "J"-like nonlinear correlation trends in some samples. In addition, competitive binding assays of the HoblOBP1 and HoblOBP2 mixtures and of the HoblOBP2 and HoblOBP4 mixtures with representative compounds unexpectedly demonstrated good affinity, which revealed extreme differences that were only obtained using the individual proteins. In the immunocytochemical analysis, colocalization of HoblOBP1 and HoblOBP2, and of HoblOBP2 and HoblOBP4, was detected in the sensilla basiconica and sensilla placodea, respectively. All of these results suggested that HoblOBP1 and HoblOBP2, as well as HoblOBP2 and HoblOBP4, may serve as heterodimers in the sensillum lymph

Effect of Oxygen Content on Microstructure and Tensile Properties of a 22Cr-5Al ODS Steel

The high tensile strength and irradiation resistance of oxide dispersion strengthened (ODS) ferritic steels is attributed to the ultrafine and dispersed oxides within the matrix. The high content of oxygen and yttrium is critical for the formation of dense Y-rich oxides. However, only few studies have reported the effect of oxygen content on the microstructure and mechanical properties of ODS steels. Herein, we employed gas atomization reactive synthesis to prepare pre-alloy powders and then hot isostatic pressing (HIP) to consolidate two 22Cr-5Al ODS steels with different oxygen content. Our results showed Y-rich precipitates at and near grain boundaries of the as-HIPed alloys. Moreover, with the oxygen content increasing from 0.04 to 0.16 wt%, more precipitates precipitated in the as-HIPed alloy, and the ultimate tensile strength of the alloy was improved. However, increasing the oxygen content to 0.16 wt% led to formation of stripe and chain precipitates at and near grain boundaries, which caused a partial intergranular fracture of the as-HIPed alloy

Functional characterization of chemosensory proteins in the scarab beetle, Holotrichia oblita Faldermann (Coleoptera: Scarabaeida).

Chemosensory proteins (CSPs) play important roles in chemical communication by insects, as they recognize and transport environmental chemical signals to receptors within sensilla. In this study, we identified HoblCSP1 and HoblCSP2 from a cDNA library of Holotrichia oblita antennae, successfully expressed them in E. coli and purified them by Ni ion affinity chromatography. We then measured the ligand-binding specificities of HoblCSP1 and HoblCSP2 to 50 selected ligands in a competitive binding assay. These results demonstrated that HoblCSP1 and HoblCSP2 have similar ligand-binding spectra. Both proteins displayed the highest affinity for β-ionone, α-ionone and cinnamaldehyde, indicating that they prefer binding to odorants other than sex pheromones. Additionally, immuno-localization revealed that HoblCSP1 is highly concentrated in sensilla basiconica, while HoblCSP2 is specifically localized to sensilla placodea. In conclusion, HoblCSP1 and HoblCSP2 are responsible for binding to general odorants with slightly different specificities due to their different in vivo environments

Effect of Thermo-Mechanical Treatment on the Microstructure and Tensile Properties of the Fe-22Cr-5Al-0.1Y Alloy

Oxide dispersion strengthened ferritic steel is considered an important structural material in fusion reactors due to its excellent resistance to radiation and oxidation. Fine and dispersed oxides can be introduced into the matrix via the powder metallurgy process. In the present study, large grain sizes and prior particle boundaries (PPBs) formed in the FeCrAlY alloy prepared via powder metallurgy. Thermo-mechanical treatment was conducted on the FeCrAlY alloy. Results showed that microstructure was optimized: the average grain diameter decreased, the PPBs disappeared, and the distribution of oxides dispersed. Both ultimate tensile strength and elongation improved, especially the average elongation increased from 0.5% to 23%

The Cinnamyl Alcohol Dehydrogenase Gene Family in Melon (<i>Cucumis melo</i> L.): Bioinformatic Analysis and Expression Patterns

<div><p>Cinnamyl alcohol dehydrogenase (CAD) is a key enzyme in lignin biosynthesis. However, little was known about CADs in melon. Five CAD-like genes were identified in the genome of melons, namely <i>CmCAD1</i> to <i>CmCAD5</i>. The signal peptides analysis and CAD proteins prediction showed no typical signal peptides were found in all <i>CmCADs</i> and <i>CmCAD</i> proteins may locate in the cytoplasm. Multiple alignments implied that some motifs may be responsible for the high specificity of these CAD proteins, and may be one of the key residues in the catalytic mechanism. The phylogenetic tree revealed seven groups of CAD and melon CAD genes fell into four main groups. <i>CmCAD1</i> and <i>CmCAD2</i> belonged to <i>the bona fide</i> CAD group, in which these CAD genes, as representative from angiosperms, were involved in lignin synthesis. Other <i>CmCADs</i> were distributed in group II, V and VII, respectively. Semi-quantitative PCR and real time qPCR revealed differential expression of <i>CmCADs</i>, and <i>CmCAD5</i> was expressed in different vegetative tissues except mature leaves, with the highest expression in flower, while <i>CmCAD2</i> and <i>CmCAD5</i> were strongly expressed in flesh during development. Promoter analysis revealed several motifs of CAD genes involved in the gene expression modulated by various hormones. Treatment of abscisic acid (ABA) elevated the expression of <i>CmCADs</i> in flesh, whereas the transcript levels of <i>CmCAD1</i> and <i>CmCAD5</i> were induced by auxin (IAA); Ethylene induced the expression of <i>CmCADs</i>, while 1-MCP repressed the effect, apart from <i>CmCAD4</i>. Taken together, these data suggested that <i>CmCAD4</i> may be a pseudogene and that all other <i>CmCADs</i> may be involved in the lignin biosynthesis induced by both abiotic and biotic stresses and in tissue-specific developmental lignification through a CAD genes family network, and <i>CmCAD2</i> may be the main CAD enzymes for lignification of melon flesh and <i>CmCAD5</i> may also function in flower development.</p></div

Activation of melanocortin-1 receptor signaling in melanoma cells impairs T cell infiltration to dampen antitumor immunity

Abstract Inhibition of T cell infiltration dampens antitumor immunity and causes resistance to immune checkpoint blockade (ICB) therapy. By in vivo CRISPR screening in B16F10 melanoma in female mice, here we report that loss of melanocortin-1 receptor (MC1R) in melanoma cells activates antitumor T cell response and overcomes resistance to ICB. Depletion of MC1R from another melanocytic melanoma model HCmel1274 also enhances ICB efficacy. By activating the GNAS-PKA axis, MC1R inhibits interferon-gamma induced CXCL9/10/11 transcription, thus impairing T cell infiltration into the tumor microenvironment. In human melanomas, high MC1R expression correlates with reduced CXCL9/10/11 expression, impaired T cell infiltration, and poor patient prognosis. Whereas MC1R activation is restricted to melanoma, GNAS activation by hotspot mutations is observed across diverse cancer types and is associated with reduced CXCL9/10/11 expression. Our study implicates MC1R as a melanoma immunotherapy target and suggests GNAS-PKA signaling as a pan-cancer oncogenic pathway inhibiting antitumor T cell response