Biochimica et biophysica acta. General subjects最新文献

{"title":"Size of aquaporin-4 orthogonal arrays of particles are affected by the palmitoylation state of the M1 isoform","authors":"Jessica D. Carder ,&nbsp;Barbara Barile ,&nbsp;Alessia Memeo ,&nbsp;Eric P. Jacobo ,&nbsp;Grazia Paola Nicchia ,&nbsp;James A. Brozik","doi":"10.1016/j.bbagen.2025.130866","DOIUrl":"10.1016/j.bbagen.2025.130866","url":null,"abstract":"<div><div>Human aquaporin-4 (AQP4) is a water-channel protein crucial for water-ion homeostasis in the central nervous system. Dysregulation of AQP4 function is linked to neurological conditions like brain edema, neuromyelitis optica, and epilepsy. AQP4 has two isoforms, M1 and M23. The M23 isoform forms large structures known as Orthogonal Arrays of Particles (OAPs), while M1 coats the outer surface of OAPs. The dynamics of OAP aggregation may modulate water permeability and ion homeostasis in the brain. This study shows that the palmitoylation state of M1 influences OAP self-assembly, with depalmitoylated M1 producing OAPs 20 % larger than those formed by palmitoylated M1 at the same concentration. This, along with prior research on M1 aggregation, suggests palmitoylated M1 forms a single outer layer arounf OPAs, whereas depalmitoylated M1 creates a double layer. Single-particle tracking was used to study M23 OAP formation in lipid bilayers under equilibrium conditions. Our data support the idea that M1 regulates M23 OAP size, showing that average OAP size decreases as M1 concentration increases. This study explores the inhibitory effects of M1 on AQP4 M23 assembly and how M1's palmitoylation state affects OAP size regulation. These insights could lead to new therapeutic approaches for managing AQP4 assembly and function in related conditions.</div></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1869 12","pages":"Article 130866"},"PeriodicalIF":2.2,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145278893","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Abscisic acid-mediated modulation of morpho-physiological traits and transcript accumulation improves drought resilience in Crocus sativus L.","authors":"Suman Gusain ,&nbsp;Rakesh Kumar ,&nbsp;Rohit Joshi","doi":"10.1016/j.bbagen.2025.130849","DOIUrl":"10.1016/j.bbagen.2025.130849","url":null,"abstract":"<div><div>Drought is a major abiotic factor leading to decreased productivity. The current study investigates the effects of prolonged drought stress and subsequent recovery in <em>Crocus sativus</em> L. through the exogenous application of abscisic acid (ABA) under field conditions in the Dhauladhar range of the Himalayan region. To evaluate the influence of drought stress, saffron corms (both large and medium) were subjected to three treatments: irrigation, water deficit (withheld irrigation), and ABA application (via foliar spray). The results indicated that morphological and physiological traits, including fresh weight and dry weight (leaf, root, mother corm and daughter corm), sprouting percentage, total biomass, flower number, stomatal aperture, relative water content, photosynthetic activity and Fv/Fm were negatively affected by drought stress. Further, histological analysis revealed a reduction in starch granule accumulation, while root metaxylem cells were found to be enlarged under water deficit conditions. Additionally, RT-PCR analysis indicated higher transcript abundance of <em>DREB1, DREB2</em> and <em>SnRK2</em> under drought conditions compared to the control, while the expression levels of <em>MYB37, WRKY1, DHN1, DHN1</em> and <em>AREB1</em> remained unchanged under similar conditions. Nevertheless, exogenous abscisic acid improved the drought tolerance of <em>C. sativus</em>. A decrease in lipid peroxidation and an increase in proline, chlorophyll content, and antioxidant activity were observed when ABA was applied under drought conditions. Overall, our study demonstrates ABA-mediated regulation of several key transcription factors under drought stress. These findings provide new insights into the mechanisms of drought tolerance in saffron, which will facilitate future breeding programmes for this highly valuable crop.</div></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1869 11","pages":"Article 130849"},"PeriodicalIF":2.2,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144879352","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a monomeric recombinant Butea monosperma agglutinin as a diagnostic and prognostic biomarker for cholangiocarcinoma","authors":"Sirintra Udomkitkosol ,&nbsp;Pondthip Thaiorn ,&nbsp;Phisit Sintusen ,&nbsp;Kulthida Vaeteewoottacharn ,&nbsp;James R. Ketudat-Cairns ,&nbsp;Sumalee Obchoei ,&nbsp;Siriporn Proungvitaya ,&nbsp;Tin May Aung ,&nbsp;Atsushi Kuno ,&nbsp;Sayaka Fuseya ,&nbsp;Atit Silsirivanit ,&nbsp;Sopit Wongkham ,&nbsp;Sukanya Luang","doi":"10.1016/j.bbagen.2025.130852","DOIUrl":"10.1016/j.bbagen.2025.130852","url":null,"abstract":"<div><div>Native <em>Butea monosperma</em> agglutinin (nBMA), is a lectin isolated from the seeds of the <em>Butea monosperma</em> plant, which binds specifically to galactose, <em>N</em>-acetylgalactosamine, and lactose. This study developed a recombinant β-chain of BMA (rBMA) expressed in <em>Escherichia coli</em>. The rBMA exists in a monomeric form, retains native structure and sugar-binding capacity without exhibiting hemagglutination activity. The binding activity of rBMA was evaluated through lectin-cytofluorescent staining of CCA cell lines. Similar to nBMA, rBMA exhibited a positive signal to CCA cell lines but displayed a strong signal in different cell lines. Sodium periodate treatment abolished rBMA binding in CCA tissues and serum dot blots, confirming carbohydrate-dependent interactions. The neutralizing activity for sugar binding specificity indicated that rBMA binds to the complex glycosylated glycans rather than mono- and di-saccharides. Elevated levels of rBMA binding glycans in serum dot blots were found to differentiate CCA patients from healthy individuals, achieving a diagnostic sensitivity of 92.9%, specificity of 36%, and overall accuracy of 74%. High levels of serum rBMA-binding glycans were associated with poorer survival in CCA patients, and directly correlated with serum alkaline phosphatase levels. No correlation was found with carcinoembryonic antigen and CA19–9 levels. These findings position serum rBMA-binding glycans as potential biomarkers reflecting CCA progression. The monomeric nature and retained glycan specificity of rBMA, coupled with its absence of hemagglutination activity, make it a superior candidate to nBMA for diagnostic applications and a promising platform for targeted therapeutic development for CCA.</div></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1869 11","pages":"Article 130852"},"PeriodicalIF":2.2,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144917164","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Unraveling the power of TOMM40: Driving PHB1-mediated mtDNA release and mitophagy to fuel breast cancer progression","authors":"Lei Pan,&nbsp;Feixia Ma,&nbsp;Hongchen Zhang,&nbsp;Yin Duan","doi":"10.1016/j.bbagen.2025.130851","DOIUrl":"10.1016/j.bbagen.2025.130851","url":null,"abstract":"<div><h3>Background</h3><div>In breast cancer (BRCA), mitophagy is essential for the survival and metastasis of cancer cells. However, the interaction between translocase of the outer mitochondrial membrane 40 (TOMM40) and prohibitin 1 (PHB1) in regulating mitophagy in BRCA remains poorly understood.</div></div><div><h3>Methods</h3><div>Based on bioinformatics analysis, the interaction between PHB1 and key mitophagy regulators in BRCA was explored. The effects of mitochondrial division inhibitor-1 (Mdivi-1) and Fluorizoline on mitophagy, cell viability, and sphere formation ability in MDA-MB-231 cells were assessed. In the cell model activated by carbonyl cyanide <em>m</em>-chlorophenylhydrazone (CCCP) to induce mitophagy, the effects of TOMM40 on cell viability, sphere formation ability, mitochondrial membrane potential, reactive oxygen species (ROS) levels, mitochondrial DNA (mtDNA) release, and PHB1 regulation were analyzed. In vivo, the impact of TOMM40 knockdown on tumor progression and mitophagy was also evaluated.</div></div><div><h3>Results</h3><div>PHB1 interacted with TOMM40. Mdivi-1 or Fluorizoline treatment inhibited mitophagy, and significantly reduced BRCA cell viability and sphere formation. CCCP treatment induced mitophagy, increased mtDNA release and PHB1 levels, decreased mitochondrial membrane potential and ROS, and promoted cell viability and sphere formation ability, which were all reversed by TOMM40 knockdown. Additionally, TOMM40 knockdown led to decreased PHB1 levels and increased ROS accumulation in tumor tissue, thus repressing tumor progression.</div></div><div><h3>Conclusion</h3><div>This study identifies TOMM40 as a key regulator that enhances PHB1-mediated mtDNA release and induces mitophagy in BRCA cells, thus promoting breast cancer progression.</div></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1869 11","pages":"Article 130851"},"PeriodicalIF":2.2,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144858810","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Inhibition of phosphodiesterase 4 and 7 regulates breast cancer cell proliferation","authors":"Carla Mayora Justel ,&nbsp;Tamara Valladares ,&nbsp;Lucía Gargiulo ,&nbsp;Verónica González-Pardo ,&nbsp;Maximiliano De Sousa ,&nbsp;María del Carmen Esandi ,&nbsp;Carlos Davio ,&nbsp;Isabel Lüthy ,&nbsp;Ariana Bruzzone","doi":"10.1016/j.bbagen.2025.130850","DOIUrl":"10.1016/j.bbagen.2025.130850","url":null,"abstract":"<div><h3>Purpose</h3><div>cAMP regulates key processes in mammary cell biology. Previous studies suggested reduced cAMP production in more malignant cells. This study investigates the role of cAMP in mammary biology using non-tumor (MCF-10A  and HBL-100) and tumor (MCF-7 and MDA-MB-231) human breast cell lines.</div></div><div><h3>Methods</h3><div>cAMP levels were quantified using a competitive radio-binding assay. Cell proliferation and viability were assessed by cell counting and MTT assay. Gene expression was analyzed by real-time PCR and immunofluorescence. Additional assays included migration, colony formation, annexin V/IP staining, comet assay, and caspase-3 activity. Public datasets were consulted. Phosphodiesterase (PDE) inhibitors were tested: the broad-spectrum PDE inhibitor IBMX (3-Isobutyl-1-methylxanthine), the PDE4-selective inhibitor roflumilast, and the PDE7-selective inhibitor BRL-50481.</div></div><div><h3>Results</h3><div>Non-tumor cells produced more cAMP than tumor cells, with or without IBMX. IBMX decreases cell proliferation and viability in all cell lines. Gene expression data revealed higher ADCY2, 3, 4, 5, 6, and 8 expression in normal tissues. Roflumilast reduced cell viability in all tested cells, while the PDE7-specific inhibitor BRL-50481 only affected MCF-7 cells. All PDE inhibitors exhibited an additive effect with tamoxifen, reducing MCF-7 cell viability. In tumor cells roflumilast decreased cell migration. In MDA-MB-231 cells, although IBMX and roflumilast showed a trend toward further decreasing viability compared to doxorubicin or paclitaxel alone, the differences were not statistically significant.</div></div><div><h3>Conclusion</h3><div>The selective PDE4 inhibitor roflumilast demonstrated potential as a therapeutic agent when combined with specific breast cancer treatments, offering a novel approach in breast cancer therapy.</div></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1869 11","pages":"Article 130850"},"PeriodicalIF":2.2,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144858809","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Hik2-Rre1 interaction acts as a two-component signaling system in filamentous cyanobacterium Anabaena PCC 7120","authors":"Priyanka Pathak ,&nbsp;Manisha Banerjee","doi":"10.1016/j.bbagen.2025.130853","DOIUrl":"10.1016/j.bbagen.2025.130853","url":null,"abstract":"<div><div>Photosynthetic organisms often rely on two-component regulatory system to adapt to environmental changes. This system is crucial for connecting external signals with the response mechanism by controlling gene expression, eventually allowing the organism to acclimatize to various stresses. Cyanobacteria, in particular, possess a large number of these two-component systems. Chloroplast Sensor Kinase (CSK) is a conserved histidine kinase present in all photosynthetic organisms.</div><div>In the present study, Hik2, a CSK homolog found in the filamentous cyanobacterium <em>Anabaena</em> PCC 7120, was investigated to understand its role in the signaling mechanism of this organism. Recombinant Hik2 was found to undergo autophosphorylation on a conserved histidine residue, which remains unaffected by low salt concentrations but is slightly inhibited at elevated concentrations. Rre1 was identified as a potential partner for Hik2 through <em>in silico</em> analysis. Further experiments, including pull-down and surface plasmon resonance analysis, confirmed a physical interaction between Hik2 and Rre1. Interestingly, rapid dephosphorylation of Hik2 in the presence of Rre1 suggested a phosphotransfer from Phospho-Hik2 to its cognate partner Rre1. <em>In silico</em> analysis further identified probable heat-responsive regulons of <em>Anabaena</em> Rre1, suggesting the possible role of the Hik2-Rre1 interaction in the signaling mechanism of <em>Anabaena</em> PCC 7120.</div><div>Overall, this study sheds light on the importance of the Hik2-Rre1 interaction in facilitating signaling processes in filamentous cyanobacteria, providing valuable insights into the acclimatization mechanisms of these photosynthetic organisms.</div></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1869 11","pages":"Article 130853"},"PeriodicalIF":2.2,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144940734","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhancing soybean tolerance to drought by homologous expression of cytokinin synthase gene GmIPT10","authors":"Xuan Lan Thi Hoang ,&nbsp;Nguyen Nguyen Chuong ,&nbsp;Nguyen Cao Nguyen ,&nbsp;Nguyen Ngoc Hai ,&nbsp;Dung Tien Le ,&nbsp;Yasuko Watanabe ,&nbsp;Keiichi Mochida ,&nbsp;Tien-Dung Nguyen ,&nbsp;Henry T Nguyen ,&nbsp;Lam-Son Phan Tran ,&nbsp;Nguyen Phuong Thao","doi":"10.1016/j.bbagen.2025.130848","DOIUrl":"10.1016/j.bbagen.2025.130848","url":null,"abstract":"<div><div>Climatic change-induced osmotic stresses, especially drought and salinity, have arisen as major environmental constraints to crop productivity and sustainable agriculture. Previously, soybean <em>GmIPT10</em>, which encodes an adenine isopentenyl transferase enzyme working in the biosynthesis of cytokinin phytohormone, has been identified as a drought-responsive gene. In this study, the aim is to explore the drought-associated attributes of GmIPT10 <em>in planta</em>, by using homologous expression system. Our findings demonstrated that the transgenic plants might acquire better drought tolerance potential. Following the drought application at vegetative stage, they not only had higher drought-tolerance index by 3–4-fold but also displayed certain advantages in maintaining agronomic traits such as better plant growth, dry biomass accumulation and cellular water contents under adverse conditions than the wild-type plants. Importantly, the greater enhancement in antioxidant enzymatic activities in the transgenic plants (<em>i.e.</em> 2.4–3.8-fold increase) compared with the WT counterparts (1.2–2.3-fold increase) indicated the better defense ability towards drought-induced oxidative stress of the former group. Additional investigation on the drought effects at the reproductive stage further highlighted a less inhibition status of the photosynthetic activities in the transgenic lines, whereby they displayed more active gaseous exchange, higher chlorophyll contents and photochemical efficiency. Although there was no difference in average seed weights, the drought-treated transgenic plants could maintain higher average pod numbers by 10 %, which contributed to higher productivity. Taking these data altogether, our results demonstrated the beneficial role of soybean IPT10 and its mediating actions in alleviating the adverse drought effects on plants.</div></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1869 11","pages":"Article 130848"},"PeriodicalIF":2.2,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144841908","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Phase separation via protein-protein and protein-RNA networks coordinates ribosome assembly in the nucleolus","authors":"Priyanka Dogra ,&nbsp;Richard W. Kriwacki","doi":"10.1016/j.bbagen.2025.130835","DOIUrl":"10.1016/j.bbagen.2025.130835","url":null,"abstract":"<div><div>The nucleolus, the largest membraneless organelle in the nucleus, functions as the site for ribosome biogenesis. While long known for its complex and dynamic structure, our mechanistic understanding of nucleolar organization has advanced dramatically in the past 15 years. The process of phase separation (PS) facilitates the compartmentalization of ribosomal components with assembly factors in the nucleolus, underlying complex ribosome biogenesis processes. Multicomponent PS creates multiple nucleolar sub-compartments that function from inside out as a ribosome assembly line. In this review, we discuss the molecular basis of nucleolar organization, including how different types of protein-protein and protein-RNA interactions create the multilayered architecture that enables ribosome biogenesis. Key proteins including nucleolin, fibrillarin, and nucleophosmin mediate nucleolar compartmentalization through their unique structural features and multivalent interactions. The processes of ribosomal RNA (rRNA) transcription, modification and splicing, and folding are spatially and temporally segregated within different regions of the nucleolus. rRNA matures and changes form along this processing continuum, continually altering its interactions with proteins, creating multiple separate liquid phases that establish sub-compartments. We highlight how both folded domains and intrinsically disordered regions (IDRs) in nucleolar proteins contribute to multivalent interactions underlying PS and nucleolar compartmentalization. We also discuss how perturbation of nucleolar PS alters nucleolar structure, dynamics, and function and contributes to a range of pathological conditions.</div></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1869 10","pages":"Article 130835"},"PeriodicalIF":2.8,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144633262","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"MiR-2779-x, a key microRNA that is related to the tumorigenicity of the MDCK cell line","authors":"Di Yang ,&nbsp;Lingwei Huang ,&nbsp;Jiachen Shi ,&nbsp;Zhenbin Liu ,&nbsp;Jiamin Wang ,&nbsp;Zhongren Ma ,&nbsp;Ayimuguli Abudureyimu ,&nbsp;Zilin Qiao ,&nbsp;Jianguo Chen","doi":"10.1016/j.bbagen.2025.130843","DOIUrl":"10.1016/j.bbagen.2025.130843","url":null,"abstract":"<div><div>The tumorigenicity of MDCK cell line is a major concern with respect to its safety for vaccine production, the effect of miRNAs on the tumorigenicity of MDCK cells is poorly understood. In this study, we performed miRNA-Seq on two MDCK cell lines with tumorigenic potential and their derived monoclonal cell lines that lack tumorigenicity. Through bioinformatics analysis, we identified differentially expressed miRNAs and conducted GO and KEGG pathway analyses of their target genes. Our results indicated that miR-2779-x and its target genes exhibited the most significant characteristics associated with tumorigenesis. Injection of live cells overexpressing miR-2779-x into nude mice resulted in a markedly reduced tumorigenesis rate (1/10). Overexpression of miR-2779-x significantly decreased the proliferation and migration capabilities of MDCK cells while enhancing their invasive potential. To identify and localize miR-2779-x target genes, we employed bioinformatics prediction, RT-qPCR, immunofluorescence assays (IFA), fluorescence in situ hybridization (FISH), and dual-luciferase reporter assays. We found that miR-2779-x negatively regulates the expression of PI3KR1 and Caspase 9 at both the gene and protein levels. Additionally, miR-2779-x co-localizes with PI3KR1 in the cytoplasm and directly targets the 3’UTR of PI3KR1. Overexpression of PI3KR1 in miR-2779-x-overexpressing cells restored cellular functions, leading to increased proliferation and migration but decreased invasion. Moreover, miR-2779-x modulates MDCK cell growth and tumorigenesis by influencing the expression and phosphorylation levels of proteins involved in the PI3K/AKT and apoptosis signaling pathways. We proposed the key pathways of miRNA involvement in MDCK cell tumorigenicity and initially revealed the function of miR-2779-x in MDCK cell tumorigenicity.</div></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1869 10","pages":"Article 130843"},"PeriodicalIF":2.2,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144673880","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Coupling of muscarinic receptors to protein kinase C underlies a feedback regulation of cell responsiveness to acetylcholine","authors":"Ekaterina A. Dymova,&nbsp;Olga A. Rogachevskaja,&nbsp;Vladislav V. Sokolov,&nbsp;Elizaveta Е. Kopylova,&nbsp;Natalia V. Kabanova,&nbsp;Stanislav S. Kolesnikov","doi":"10.1016/j.bbagen.2025.130844","DOIUrl":"10.1016/j.bbagen.2025.130844","url":null,"abstract":"<div><div>Acetylcholine (ACh)-induced Ca²⁺ signaling was analyzed in HEK-293 (WT-HEK) cells and their derivatives, IP3R1-HEK, IP3R2-HEK, and IP3R3-HEK with a single functional IP₃ receptor isoform, IP₃R1, IP₃R2, or IP₃R3, respectively. The initial stimulation of WT-HEK cells triggered a prolonged feedback process that diminished their responsiveness to ACh. Inhibition of protein kinase C (PKC) with Gö 6983 or calphostin C prevented the decline of ACh responsivity, indicating that PKC was involved. Using IP3R1-HEK, IP3R2-HEK, and IP3R3-HEK cells, it was shown that PKC was capable of regulating Ca²⁺ release through each IP₃R isoform. While in control, IP₃ uncaging triggered Ca²⁺ transients in ∼15 % of cells loaded with caged-Ins(145)P3/PM, PKC inhibition enlarged this fraction nearly twofold. These observations suggested that in ACh transduction machinery, PKC targeted primarily IP₃-driven Ca²⁺ release. ADP and 5-HT triggered Ca²⁺ transients in WT-HEK cells and CHO cells expressing endogenous P2Y and recombinant 5HT2C receptors, respectively. The responsiveness of WT-HEK cells to ADP and CHO cells to 5-HT applied serially declined after the initial cell stimulation but PKC inhibition precluded this phenomenon almost completely. The coupling of GPCRs to PKC in living cells, muscarinic and P2Y receptors in WT-HEK cells and 5HT2C receptors in CHO cells, was demonstrated for the first time using real-time fluorescence imaging and sapphireCKAR, a genetically encoded sensor of PKC activity. Altogether, our findings suggest that a PKC-based feedback regulation of agonist-induced Ca²⁺ release might be a common attribute of transduction of various agonists involving GPCRs coupled to the phosphoinositide cascade.</div></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1869 10","pages":"Article 130844"},"PeriodicalIF":2.2,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144717301","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}