Waterlogging is a major abiotic stress that severely limits grapevine productivity by inducing hypoxia and oxidative stress in the root tissues. Transcription factors, such as APETALA2/Ethylene-Responsive Factor (AP2/ERF) are closely involved in regulating plant survival under waterlogging stress. Here, we characterized VvERF113, a root-enriched AP2/ERF transcription factor from Vitis vinifera, and investigated its role in regulating plant waterlogging tolerance. VvERF113 overexpression in Arabidopsis thaliana significantly alleviated leaf wilting and oxidative damage under waterlogged conditions, accompanied by increased antioxidant enzyme activity and reduced hydrogen peroxide accumulation. Molecular analyses revealed that VvERF113 enhanced the expression of hypoxia-responsive genes (AtADH1 and AtPDC1), ethylene-related HD-Zip transcription factors (AtHB1 and AtHB2), and oxygen-sensing genes (AtPCO1 and AtPCO2), indicating its involvement in multiple regulatory pathways. VvERF113 physically interacts with VvATHB-13, forming a transcriptional module that integrates AP2/ERF and HD-Zip I signaling. This interaction was validated using yeast two-hybrid, bimolecular fluorescence complementation, and glutathione S-Transferase pull-down assays. This study identified a novel ERF-HD-ZIP regulatory node that coordinates fermentative metabolism, ethylene signaling, and redox balance under hypoxic stress. Conclusively, VvERF113 presents a promising candidate for breeding waterlogging-tolerant grapevine cultivars.

BACKGROUND: Anterior Cruciate Ligament (ACL) reconstruction using a triple hamstring semitendinosus graft is a commonly used technique for optimal hamstring function, flexion strength, and rapid recovery. In the conventional method, the end of the tendon graft is whipstitched to a suspensory loop, which reportedly can lead to graft failure due to tendon laceration or slippage. This study aims to enhance ACL fixation by introducing a novel implant device replacing conventional sutures. METHODS: Six initial designs were introduced, and a product design specification (PDS) chart was used to select one. The design named (Zip-Tie) was chosen based on the PDS scoring and three variants of it were prototyped and subjected to in-vitro experiments to optimize the design. The best performing variant was chosen as the final design which underwent additional validation tests. The mechanical experiments consisted of three loading steps, a preconditioning, a main cyclic, and a pull-out loading. RESULTS: The mechanical properties of the three device variants were compared, and the best performing one was selected as the final design. The final design exhibited superior mechanical properties compared to similar studies, with an average cyclic stiffness (ACS) of 37,637 ± 8,910 N/mm, average pull-out stiffness (APS) of 132.8 ± 28.9 N/mm, and cyclic elongation of 1.11 ± 0.27 mm. The load-to-failure results showed that 80% of the samples exceeded 1000 N. CONCLUSIONS: The introduced novel implant device for preparing tripled semitendinosus grafts in ACL reconstruction, demonstrated superior mechanical performance compared to conventional suturing methods. The integration of friction plates and straps enhanced graft fixation and stability. These results support the potential of a new fixation approach, laying the groundwork for future in vivo studies and the exploration of optimal biocompatible materials for clinical application.

Drought represents a major abiotic factor that restricts crop yields worldwide. Previous studies have demonstrated that the wheat bZIP transcription factor TabZIP156 plays a vital role in enhancing drought tolerance. However, the drought tolerance mechanism of TabZIP156 in wheat remains unclear. In this study, TaJAZ3-2A was identified as an interactor of TabZIP156 through yeast two-hybrid screening. This interaction was confirmed in vivo using luciferase complementation imaging (LCI) and bimolecular fluorescence complementation, and further validated in vitro by GST pull-down experiments. The LCI assays showed that the C-terminal domain of TabZIP156 interacted with the N-terminal domain of TaJAZ3-2A. Transcription level analysis revealed that TaJAZ3-2A was predominantly expressed in roots, and exhibited significant response to PEG 6000-induced drought, ABA, and MeJA stress. Subcellular localization analysis showed that TaJAZ3-2A was localized to the nucleus. Functional studies revealed that TaJAZ3-2A overexpressing Arabidopsis displayed reduced drought tolerance. Conversely, virus-induced gene silencing of TaJAZ3-2A in wheat showed enhanced drought tolerance and upregulated key stress-responsive and antioxidant-related genes. Moreover, the dual-luciferase assays showed that TaJAZ3-2A and TabZIP156 mutually antagonize each other in regulating the expression levels of TaP5CS, TaDREB1A, and TaPOD. In conclusion, these findings provide novel mechanistic insights into the role of TabZIP156 in wheat drought tolerance and establish a theoretical foundation for the development of drought-tolerant wheat cultivars.

Rationale: Both bulk RNA-sequencing and GEO database upon chemotherapy to non-small cell lung cancer (NSCLC) cells reveal that ZNF131 (Zinc Finger Protein 131) maybe a crucial transcriptional factor involved. However, it is a recently discovered protein with largely unexplored expression patterns and biological functions. Methods: Bioinformatics analyses and immunohistochemistry staining were assessed to detect both mRNA and protein levels of ZNF131 in NSCLC specimens and cell lines. Next, colony formation assay, MTT assay, EdU assay, transwell assay, flow cytometric analysis, sphere formation assay, western blotting analysis, mouse xenograft model analysis, immunofluorescence assay, and reverse transcriptase-polymerase chain reaction were performed to investigate the effect of ZNF131 interaction on proliferation, invasion, stemness, chemotherapy sensitivity. RNA-sequencing assay, RNA-microarray, and ChIP-sequencing assay were used to identify candidate downstream target genes. Further, liquid chromatography-tandem mass spectrometry analysis, GST pull-down assay, and immunoprecipitation assays were performed to evaluate the interactions between ZNF131, BACH1, and CUL3. Results: ZNF131 was elevated in NSCLC specimens and cell lines, which significantly correlates with advanced TNM stage and poor prognosis in NSCLC patients. ZNF131 overexpression promotes NSCLC cell proliferation, invasion, and stemness both in vitro and in vivo. ZNF131 appears to target the RAD51 gene within a well-defined region (-668bp to -403bp) of the RAD51 promoter. ZNF131 contributes to RAD51-dependent homologous recombination (HR), primarily through its Zinc Finger and BTB domains. ZNF131-BACH1 interaction, mediated by their respective BTB domains, enhances the stability of both proteins, effectively preventing their ubiquitin-mediated degradation by CUL3. The ZNF131-BACH1 partnership significantly amplifies RAD51-dependent HR, resulting in expedited resistance to both radiotherapy and chemotherapy in NSCLC patients. Desoxyrhaponticin was shown to halt NSCLC progression and orchestrate a synergistic effect together with chemotherapy at least partially by targeting ZNF131. Conclusions: Our findings indicate that ZNF131 exhibits heightened expression in NSCLC, driving essential processes such as proliferation, invasion, and stemness by transcriptionally activating RAD51. The ZNF131-BACH1 interaction serves as a crucial enhancer, further boosting RAD51 transcription and ultimately accelerating therapy resistance in NSCLC.

Sixteen patient-derived xenografts (PDXs) were analyzed using a mass spectrometry (MS)-based kinase inhibitor pull-down assay (KIPA), leading to the observation that death-associated protein kinase 3 (DAPK3) is significantly and specifically overexpressed in the triple-negative breast cancer (TNBC) models. Validation studies confirmed enrichment of DAPK3 protein, in both TNBC cell lines and tumors, independent of mRNA levels. Genomic knockout of DAPK3 in TNBC cell lines inhibited in vitro migration and invasion, along with down-regulation of an epithelial-mesenchymal transition (EMT) signature, which was confirmed in vivo. The kinase and leucine-zipper domains within DAPK3 were shown by a mutational analysis to be essential for functionality. Notably, DAPK3 was found to inhibit the levels of desmoplakin (DSP), a crucial component of the desmosome complex, thereby explaining the observed migration and invasion effects. Further exploration with immunoprecipitation-mass spectrometry (IP-MS) identified that leucine-zipper protein 1 (LUZP1) is a preferential binding partner of DAPK3. LUZP1 engages in a leucine-zipper domain-mediated interaction that protects DAPK3 from proteasomal degradation. Thus, the DAPK3/LUZP1 heterodimer emerges as a newly discovered regulator of EMT/desmosome components that promote TNBC cell migration.

Despite advancements in cancer research, the prognostic implications of competing endogenous RNA (ceRNA) networks in prostate cancer (PCa) remain incompletely understood. This study aimed to elucidate the prognostic relevance of ceRNA networks in PCa, utilizing a comprehensive bioinformatics approach alongside experimental validation. After searching The Cancer Genome Atlas (TCGA) database, RNA sequencing (RNA-Seq) data were extracted to identify differentially expressed RNAs (DERs) between 491 PCa samples and 51 normal prostate tissues, following which a comprehensive bioinformatics strategy was implemented to construct a ceRNA network. An optimal prognostic signature comprising these DERs was then established and validated using TCGA data. In addition, functional validation was performed through RNA pull-down, dual-luciferase reporter assays, quantitative real-time PCR, and western blot analysis conducted in PC-3 and DU145 cell lines, thereby complementing the bioinformatics analysis. A total of 613 DERs, comprising 103 long noncoding RNAs (lncRNAs), 60 microRNAs (miRNAs), and 450 messenger RNAs (mRNAs), were identified and utilized in constructing a ceRNA network, which encompassed 23 lncRNAs, 9 miRNAs, and 52 mRNAs. An optimal prognostic signature was established, including VPS9D1 antisense RNA 1 (VPS9D1-AS1), miR-449a, cyclin-dependent kinase 5 regulatory subunit 1 (CDK5R1), targeting protein for Xklp2 (TPX2), solute carrier family 7 member 11 (SLC7A11), copine7 (CPNE7), and maternal embryonic leucine zipper kinase (MELK), yielding area under the curve (AUC) values exceeding 0.8 across training, validation, and entire datasets. Our experiments results revealed an interaction between lncRNA TRHDE antisense RNA 1 (TRHDE-AS1) and miR-449a and that miR-449a could target the ADAM metallopeptidase with thrombospondin type 1 motif 5 (ADAMTS5) mRNA. Knockdown of miR-449a significantly impeded cell proliferation, G1/S transition, migration and invasion, and promoted apoptosis in PC-3 and DU145 cells. Furthermore, knockdown of miR-449a notably downregulated protein expression of CDK4, cyclin D1, N-cadherin and vimentin, while upregulating protein expression of cleaved caspase-3 and E-cadherin. This study contributes to a deeper understanding of the prognostic-linked ceRNA network in PCa, providing fundamental insights that could improve diagnostic and therapeutic approaches for PCa management.