MicroRNAs (miRNAs) are emerging as crucial regulators of skeletal muscle development and regeneration; however, the biological functions of many miRNAs remain to be elucidated. In this study, we focused on the function of miR-379-5p, a miRNA we previously identified as highly expressed in the longissimus dorsi muscle of goats. Overexpression of miR-379-5p inhibited the proliferation and differentiation of goat skeletal muscle satellite cells (MuSCs), as evidenced by decreased expression of proliferation and differentiation markers, reduced EdU + cells, and lower myotube formation. Through bioinformatics prediction and experimental validation, we identified LIN28B as a direct downstream target of miR-379-5p. Functional assays revealed that LIN28B promoted the proliferation and differentiation of MuSCs, whereas miR-379-5p suppressed these processes by decreasing LIN28B expression. Furthermore, miR-379-5p inhibited mitochondrial activity during the proliferation phase but promoted it during myogenic differentiation. Additionally, ectopic expression of LIN28B decreased mitochondrial membrane potential and enhanced reactive oxygen species (ROS) production, suggesting that LIN28B impairs mitochondrial function. Overall, our findings highlight the role of miR-379-5p and LIN28B in regulating goat MuSCs activity and mitochondrial function, providing new insights into the role of miRNAs in skeletal muscle development.

Sprouting, an important process in plant growth and development, is economically critical for the tea plant (Camellia sinensis (L.) O. Kuntze) and other horticultural crops, in which tender spring shoots constitute the primary harvest. While gibberellin's (GA) role in plant growth is well-established, its molecular control over bud sprouting and the underlying regulatory network remain incompletely characterized. This study employed electrophoretic mobility shift assay, dual-luciferase reporter assays, yeast one-/two-hybrid, and in situ hybridization to elucidate the transcriptional regulatory network among CsHDZ34, CsZHD9, CsDELLA5, and CsMADS27, while the functions of CsHDZ34 and CsDELLA5 in tea bud sprouting were investigated using western blotting, antisense oligodeoxynucleotides, gene overexpression, and data-independent acquisition quantitative proteomics, along with genome-wide association study analysis and high-throughput mutation detection to characterize their genetic variations. This study identified CsHDZ34 as an upstream regulator of CsZHD9, which promotes tea bud sprouting, and found that GA promoted alternative splicing in the 5' untranslated region of CsHDZ34, increasing its protein abundance in tea buds. The positive role of CsHDZ34 in bud break regulation was validated by ectopic expression in poplars and gene silencing in tea plants. CsDELLA5 was identified as being directly downstream of CsHDZ34 and as an interaction factor of CsMADS27. Notably, the CsDELLA5Hap2 variant was identified as less susceptible to GA-mediated degradation, allowing it to accumulate and act as a transcriptional activator that interacts with CsMADS27, thereby promoting bud sprouting through direct regulation of downstream target genes. These findings reveal a novel molecular mechanism orchestrated by CsHDZ34 that promotes bud sprouting in tea plants through spliceosome modulation and GA signaling. This provides a theoretical framework for elucidating bud break mechanisms and advancing early cultivar breeding.

Targeting Skp2 by camptothecin induces p27 accumulation and confers drug resistance in non-small cell lung cancer.

Fat body specific ABCG3 accounts for xenobiotic tolerance and is regulated by the homeobox transcription factor Bcd in Spodoptera litura.

Multigene engineering in plants: Technologies, applications, and future prospects.

Genome-wide identification and characterization of TCP family in apple (Malus domestica Borkh.) and functional analysis of MdTCP21 in shoot branching.

Unveiling the functional contribution of Malpighian tubule specific glucose transporters to xenobiotics tolerance in Spodoptera litura.

MYB20, an R2R3-type MYB transcription factor, negatively regulates salt and cold stress tolerance in pears.

Drought is a key limiting factor for growth of plant and crop production. In an effort to produce drought tolerant tobacco, an expression cassette comprising the Arabidopsis DREB1A cDNA under the Figwort Mosaic Virus (FMV) and Rice SalT promoters were transformed into tobacco via Agrobacterium mediated transformation. FMV is a strong and constitutive promoter and SalT is a stress inducible promoter that can be used for enhancing expression of AtDREB1A gene in tobacco. PCR was used to confirm the putative transgenic T0 plants and copy number was determined by Southern blot hybridization. RT-PCR confirmed the gene expression in transgenic tobacco lines. The selected transgenes with one copy number were subjected to osmotic stress. Seed germination results showed that transgenic seeds were able to germinate on 300 mM mannitol while control seeds were unable to germinate. Transgenic plants exhibited higher drought tolerance and produced more seeds than control plants when water was withheld for 10 days. Various physiological tests revealed higher drought stress tolerance in transgenic plants than their wild type counterparts. The current study showed that overexpression of AtDREB1A gene under FMV and SalT promoters increases drought tolerance in transgenic tobacco and offers applications in developing drought tolerant crops.

Cisplatin resistance causes ineffectiveness of cisplatin-based treatment for cervical carcinoma. The combination of cisplatin and other chemotherapeutic drugs is an available strategy to overcome this problem. However, chemotherapeutic drugs combined with cisplatin may show tissue toxicity and systemic side effects. Thus, there is a great need of seeking effective substitutes for these chemotherapeutic drugs to improve combination therapy. Here, we found that inactivating IL-6/JAK2/STAT3 signaling pathway sensitized carcinoma cells to cisplatin toxicity by increasing cisplatin accumulation, impairing DNA damage repair, and inhibiting the initiation and development of autophagy, which subsequently caused the increases in DNA damage levels and apoptosis rates in cisplatin-treated cells. We predicted that TFF3 negatively regulated transduction in the IL-6/JAK2/STAT3 pathway based on in silico analysis of the differentially expressed genes (DEGs) between highly trefoil factor 3(TFF3)-encoding mRNA-expressing carcinoma tissues and low-expressing counterparts, and experimentally determined that both ectopic expression of TFF3-encoding gene and TFF3 administration inhibited IL-6-induced STAT3 activation in carcinoma cells. Mechanistically, upon binding to IGF2R, TFF3 stabilized IGF2R by inhibiting the ubiquitin-proteasome degradation pathway to inactivate Akt and thereby STAT3. Moreover, we discovered that TFF3 administration antagonized protective effects of IL-6 stimulation against tumor-killing capacity of cisplatin. Based on these findings, we consider that TFF3 may be employed as a cisplatin sensitizer and have advantages over traditional chemotherapeutic drugs in cisplatin-based combination therapy, since it is a naturally occurring protein in cervical tissue.

Lysin motif (LysM) effectors contribute to the virulence of many pathogens, but the underlying mechanism of this type of effector remains poorly understood. Here, we identified a LysM-containing protein, named LtLysM2, in the plant opportunistic pathogen Lasiodiplodia theobromae. We demonstrated that LtLysM2 contributes to the virulence of the pathogen, is able to bind to chitin oligosaccharides, and can suppress chitin-triggered plant immune responses. Importantly, we showed that the grapevine (Vitis vinifera) protein VvSrc2, a membrane- and nuclear-localized protein homologous to the soybean (Glycine max) protein Src2 (Soybean genes regulated by cold 2), interacts with LtLysM2. Interestingly, the nuclear accumulation of VvSrc2 was elevated in the presence of LtLysM2. Ectopic expression of VvSrc2 in Nicotiana benthamiana enhanced resistance to L. theobromae. Additionally, the VvSrc2 protein interacted with the nuclear-localized RNA-binding protein VvUbp1, contributing to the programmed cell death evoked by VvUbp1. Our findings reveal a previously uncharacterized regulatory pathway in which VvSrc2 is up-regulated after the recognition of LtLysM2 and serves as an intermediator to transduce extracellular signaling events into nuclear components during infection by L. theobromae through association with the downstream target VvUbp1 in the cell nucleus.

Fbxo45 promotes cell viability, invasion and sunitinib resistance of clear cell renal cell carcinoma by targeting Erbin.

JrWOX5 promotes adventitious root formation and modulates plant architecture by interacting with key developmental regulators, providing novel insights into WOX-mediated organogenesis in woody plants. WUSCHEL-related homeobox (WOX) transcription factors, a plant-specific gene family, play essential roles in regulating plant development, including stem cell maintenance and organogenesis. Among the WOX genes identified in Juglans regia, JrWOX5 exhibited significantly elevated expression during AR formation, suggesting a potential regulatory role in this process. To investigate its function, we employed a combination of bioinformatics analysis, subcellular localization, heterologous overexpression, yeast two-hybrid (Y2H) assays, and bimolecular fluorescence complementation (BiFC). The JrWOX5 protein was localized to the nucleus. Ectopic expression of JrWOX5 in transgenic poplar markedly promoted AR formation and altered plant architecture, characterized by increased lateral branching and reduced plant height. To elucidate the molecular mechanism, protein-protein interaction (PPI) network analysis was conducted, and key candidate interactors were experimentally validated. JrWOX5 was found to interact with JrLBD16, a LOB domain-containing protein; JrLHW, a transcription factor; and JrCNR8, a regulator of cell proliferation. These findings indicate that JrWOX5 interacts with developmental regulators, which may jointly affect organogenesis and plant architectural patterning. This study aims to provide a new insight into WOX-associated organogenesis in woody plants.

Plant pathogenesis-related 1 (PR1) proteins are members of the CAP protein superfamily and are widely used immune markers. The precise function and mode of action of CAP proteins, however, remains poorly understood. In Cytospora chrysosperma, the causative agent of poplar canker disease, three CAP family members, CcCAP1-3, have been identified. CcCAP1 is required for fungal virulence and suppresses the plant immune response to facilitate fungal colonisation, in a nuclear-localisation dependent way. In this study, we functionally characterised the other two members of the C. chrysosperma CAP family, CcCAP2 and CcCAP3. Targeted gene deletions revealed that CcCAP2 and CcCAP3 affected vegetative growth and CcCAP2 contributed to fungal virulence during C. chrysosperma- Populus euramericana interaction. While CcCAP3 is predicted to be GPI-anchored, CcCAP2 is a secreted glycoprotein. Ectopic expression of CcCAP2 in Nicotiana benthamiana promoted Botrytis cinerea infection and inhibited plant immune-related gene expression. To elucidate the mode of action of the two virulence-associated proteins, CcCAP1 and CcCAP2, we employed a yeast two-hybrid-based approach and identified interacting proteins. Screening of a poplar library revealed the highest number of hits with RNA polymerase II, subunit 11 (RPB11). The interactions between CcCAP1, 2 and RPB11 were validated by colocalization analysis, FRET-FLIM, and co-immunoprecipitation analysis. Interestingly, virus-induced gene silencing of RPB11 resulted in a dwarf phenotype of the host plant, reduced fungal colonisation, and highly induced PR1 gene expression. These results suggest that the fungal virulence-associated CAP family members, CcCAP1 and CcCAP2, both target plant RNA polymerase II subunit 11 to suppress host immunity and facilitate fungal infection. The dwarf phenotype and induced expression of PR1 upon downregulation of RPB11 suggest that its function strongly affects the plant's growth-defence trade-off. These observations deepen our understanding of the complex functions of CAP family proteins in plant immune resistance and fungal virulence.

Zebrafish pou3f3b controls saccular/auditory development and marks non-neuronal cells that delaminate from the otic vesicle to promote neuroblast maturation.

Mutations in the human patatin-like lysophospholipase domain containing the 6 gene PNPLA6 encode an evolutionarily conserved (lyso)phospholipase, leading to the development of a complex hereditary spastic paraplegia 39 (SPG 39) and a number of rare severe syndromes in humans. Diseases disrupt the functioning of the nervous and reproductive systems and the gastrointestinal tract. The study aims to investigate the role of the Drosophila melanogaster swiss cheese gene, an ortholog of the human PNPLA6 gene, in gut function. We showed that the swiss cheese gene knockout leads to changes in the morphology of the midgut, disruption of the septate junction structure and the intestinal barrier permeability, and a decrease in the lipid droplet number in enterocytes. As a result of such disturbances, intestinal stem cells (ISCs) proliferation is activated, and the gut microbiome is altered. Ectopic expression of human PNPLA6 leads to the recovery of the intestinal barrier in the fly gut. The example of Drosophila demonstrates the important role of evolutionarily conserved (lyso)phospholipase in intestinal homeostasis.

ABSTRACTTranscription factor (TF) expression and dosage regulate developmental cell fate decisions. Increased TF dosage has been predicted to enhance expression of high-affinity target genes but also increase the binding of lower-affinity loci. The relative importance of high- versus lower-affinity TF binding in guiding cell fate decisions remains unclear.To test the roles of TF dosage, we examined the effects of increasing the dosage of MyoD1, the “master regulator of myogenesis”, on skeletal muscle differentiation. Unexpectedly, increased MyoD1 dosage inhibited canonical myogenesis and redirected myoblast differentiation towards forming spontaneously contracting myotubes. This novel phenotype was driven by the MyoD1-dose-dependent upregulation of non-myogenic genes, including cell adhesion genes whose ectopic expression also inhibited classical myogenic differentiation and enabled myotube contraction.Live-cell single-molecule imaging showed that elevated MyoD1 dosage increased total chromatin binding and CUT&RUN profiling demonstrated that this increase occurred via preferential binding to lower-affinity loci. Integration of CUT&RUN, ATAC-seq and RNA-seq experiments revealed that increased MyoD1 binding correlated to the upregulation of otherwise lowly expressed genes. These findings suggest that increased MyoD1 dosage induced a selective gene regulatory expansion from high- to lower-affinity cis-regulatory elements, activating a broader ensemble of target genes, revealing a TF dose-dependent mechanism that can trigger distinct developmental programs.

Growth regulatory factors (GRFs) are sequence-specific DNA-binding transcription factors that play pivotal roles in regulating plant growth and development, and in enhancing plant tolerance to biotic and abiotic stresses. Although genome-wide structural and evolutionary studies have mapped and analyzed GRF genes in different plant species, knowledge of their characteristics and functions in sunflower (Helianthus annuus) remains limited. In this study, we used bioinformatics analyses and transgenic experiments to systematically analyze the structure and function of these genes. A total of 17 HaGRF genes were identified and classified into four distinct clades, with members of the same clade sharing conserved exon-intron structures and domain architectures. All HaGRFs were predicted to localize to the nucleus, which was experimentally verified for HaGRF2c, HaGRF3, and HaGRF8c. Transcriptome analysis demonstrated tissue-specific expression and stress-responsive profiles among the HaGRF genes. Quantitative real-time PCR revealed that several HaGRF genes were significantly induced under polyethylene glycol and NaCl stress. Additionally, ectopic expression of HaGRF2c in Arabidopsis enhanced growth and conferred greater drought tolerance, supporting its dual functions in regulating growth and in adapting to stress. In summary, this research elucidates the evolutionary relationships, conserved structural characteristics, expression patterns, and roles of the HaGRF gene family in sunflowers. These findings not only deepen our understanding of the biological functions of GRF transcription factors in sunflowers but also provide valuable candidate genes for improving yield and stress resistance in H. annuus.

Chemoresistance remains a crucial obstacle in breast cancer therapy. The mechanisms underlying chemoresistance need to be explored urgently and in depth. Breast cancer metastasis suppressor 1 like (BRMS1L), a core component of the Sin3A–histone deacetylase (HDAC) co-repressor complex, has been reported to suppress breast cancer metastasis through epigenetically regulating the Wnt signal pathway. However, whether BRMS1L could regulate chemosensitivity has not been explored. Herein, we found that higher BRMS1L expression was significantly correlated with increased chemotherapy sensitivity and better prognosis in patients receiving neoadjuvant chemotherapy. In vitro experiments confirmed that chemoresistant breast cancer cells exhibited decreased BRMS1L expression compared to chemosensitive cells. In vivo experiments in nude mice demonstrated that BRMS1L markedly strengthened the chemotherapy effects on xenografts. RNA sequencing (RNA-seq) was performed to elucidate the molecular mechanism underlying BRMS1L-mediated chemosensitivity. Bioinformatics analysis indicated that BRMS1L promotes chemotherapy sensitivity by regulating cellular autophagy. Furthermore, chemoresistant breast cancer cells exhibited elevated autophagy levels, and ectopic expression of BRMS1L significantly suppressed protective autophagy through downregulating ATG5. Collectively, these results revealed that BRMS1L enhances chemotherapy sensitivity via inhibiting protective autophagy. To our knowledge, this is the first study that showed that reduced BRMS1L expression is associated with poor response to neoadjuvant chemotherapy and unfavorable prognosis in breast cancer patients. Our findings reveal a novel role of BRMS1L in chemosensitivity and highlight its potential clinical application in the treatment of breast cancer.

The root-knot nematode (RKN) Meloidogyne chitwoodi is a threat for potato production in the western United States (U.S.), negatively impacting potato yield and product value. Meloidogyne chitwoodi produce proteins, called effectors, in their esophageal glands that are secreted during parasitism and play integral roles in plant-nematode interactions. Because the esophageal glands are the main effector secretory organs, we isolated juvenile gland cells and performed gland transcriptome analysis with our newly improved genome annotation. The gland-specific transcriptome data gave us an enrichment of gland-localized genes, which was validated by in situ hybridization. The gland transcriptome analysis led to the identification of 125 effector candidates. One of the effectors that was highly expressed in the pre-parasitic J2 gland tissue, referred to as McGland26, was further characterized. Arabidopsis thaliana expressing McGland26 showed enhanced susceptibility to M. chitwoodi. However, the ectopic expression of McGland26 in planta did not suppress plant defenses, suggesting that this effector might be involved in processes other than interfering with plant immunity. Our data show that by using the gland transcriptome, a good quality genome annotation, and stringent selection criteria, we can increase the efficiency of effector identification, which can be used to develop more sustainable management tools.