Gene最新文献

{"title":"Detecting the distribution patterns of histone lactylation in the mouse testis at different developmental stages","authors":"Qian-Ni Li ,&nbsp;Fei-Chen Wang ,&nbsp;Zhen He ,&nbsp;Hai-Ping Tao ,&nbsp;Qi-En Yang","doi":"10.1016/j.gene.2025.149355","DOIUrl":"10.1016/j.gene.2025.149355","url":null,"abstract":"<div><div>Lactate is a key glycolytic metabolite that serves as an energy substance and signaling molecule. Lactylation, a recently characterized posttranslational modification (PTM), has been identified in histone and nonhistone proteins. Compelling evidence suggests that this lactate-related epigenetic modification potently regulates gene expression under physiological and pathological conditions. However, the distribution of this histone modification in the testis remains largely unknown. In this study, we investigated the expression dynamics of histone acetyltransferases (HATs), histone deacetylases (HDACs), and the lactate-regulating enzyme hexokinase 2 (HK2), and examined the cellular distribution of several types of histone lactylation, which have been identified as important for transcription and chromatin accessibility, in mouse testes during critical postnatal developmental stages. The results revealed that the expression levels of the lactylation-associated transcripts were developmentally regulated and that the histone lactylation, including H3K9la, H3K12la and H4K18la were present in spermatogenic and Sertoli cells at postnatal days (PD) 0, 6, 21, and 60. However, signals for H3K5la and H3K14la were not detected in gonocytes at PD0 and signal for H3K14la was not detected in mature Sertoli cells or spermatogonia of adult testes. Furthermore, a lack of lactate dehydrogenase a (<em>Ldha</em>) in Sertoli cells impacted the localization of several histone lactylation modifications in spermatogenic cells. Notably, H4K12la was specifically detected in zygotene and diplotene spermatocytes in the control testis, whereas it was present mainly in spermatogonia of <em>Lhda</em> Sertoli cell conditional knockout testis (<em>Ldha</em>-cKO). The results of this study lay a foundation for further understanding the role of lactylation modification in spermatogenesis and provide important data for further dissectting the role of Sertoli cell-derived lactate in germ cell development.</div></div>","PeriodicalId":12499,"journal":{"name":"Gene","volume":"948 ","pages":"Article 149355"},"PeriodicalIF":2.6,"publicationDate":"2025-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143508354","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":"Overcoming drug resistance in ovarian cancer through PI3K/AKT signaling inhibitors","authors":"Madhunika Agrawal ,&nbsp;Satyam Kumar Agrawal ,&nbsp;Kanwaljit Chopra","doi":"10.1016/j.gene.2025.149352","DOIUrl":"10.1016/j.gene.2025.149352","url":null,"abstract":"<div><div>Ovarian cancer has been identified as the eighth most typical gynaecological malignancy and cause of health problems in women. For almost forty years, platinum doublet chemotherapy has usually been the cornerstone of first-line treatment regimens. The effectiveness of conventional chemotherapy is severely hampered by relapse rates and mortality from chemotherapy resistance, which lead to the spread and recurrence of malignant cancers as well as poor outcomes in terms of quality of life in patients.</div><div>Drug resistance has been linked to several mechanisms, including increased drug efflux, decreased apoptosis, increased autophagy, and changed drug metabolism. Further, the dysregulation of tumor suppressors or oncogenes plays a crucial role in chemoresistance. Additionally, PI3K/AKT/mTOR signaling has been implicated in several <em>in vitro</em> as well as clinical studies as a significant contributor to chemotherapy resistance in case of ovarian cancer. This review discusses the potential of various crude extracts, synthetic molecules, and nanoformulations for targeting PI3K/AKT/mTOR pathway. Moreover, a range of clinical studies involving PI3K/AKT/mTOR inhibitors have been summed up, addressing both the promises and complexities associated with their use. Overall, the review aims to provide a roadmap for future investigations that could lead to improved therapeutic outcomes for patients suffering from ovarian cancer, emphasizing the urgent need for continued exploration of novel pathways and strategies to combat drug resistance.</div></div>","PeriodicalId":12499,"journal":{"name":"Gene","volume":"948 ","pages":"Article 149352"},"PeriodicalIF":2.6,"publicationDate":"2025-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143482743","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":"Disrupted maxillofacial, cardiovascular, and nervous development in washc5 knockout Zebrafish: Insights into 3C syndrome","authors":"Luyao Wei ,&nbsp;Shijun Hu ,&nbsp;Xueyang Gong ,&nbsp;Yiliya Ahemaiti ,&nbsp;Diwen Li ,&nbsp;Shi Ouyang ,&nbsp;Yuyang Huang ,&nbsp;Yongyi Wang ,&nbsp;Yan Liang ,&nbsp;Yun Deng ,&nbsp;Lin Liu ,&nbsp;Tianli Zhao","doi":"10.1016/j.gene.2025.149351","DOIUrl":"10.1016/j.gene.2025.149351","url":null,"abstract":"<div><div>3C syndrome features craniofacial, nervous, and cardiovascular malformations. <em>WASHC5</em> gene mutations may underline this syndrome, but the pathogenicity and underlying mechanism remain undetermined. We analyzed the expression pattern of the <em>washc5</em> gene in zebrafish using whole-body in situ hybridization and generated a zebrafish model with washc5 gene knockout using CRISPR/Cas9 technology. Homozygous zebrafish exhibited high mortality, retarded growth, lighter stripes, and reduced pigmentation around the pupils. In the maxillofacial region, homozygotes displayed a shortened and tilted maxilla and delayed ossification of bones. In the heart, homozygous zebrafish showed a decreased heart rate, increased ventricular area, disorganized ventricular muscle fibers, mitochondrial swelling, Golgi lysis, and endoplasmic reticulum (ER) lysis in ventricular myocytes. The mRNA levels of <em>nppb</em> and <em>myh7</em> were significantly increased. In the nervous system, homozygotes displayed bradykinesia and impaired neuronal development. qRT-PCR analysis revealed downregulation of <em>col1a2</em>, <em>col1a1a</em>, <em>col1a1b</em>, <em>sp7</em>, and <em>msx2b</em> (osteogenic factors and regulators of maxillofacial skeletal development) and abnormal expression of <em>alpk2</em>, <em>alpk3b</em>, <em>actc2</em> (cardiac development factors), as well as <em>tsen54</em>, <em>exosc8</em>, and <em>exosc9</em> (cerebellar development factors). Enrichment analysis of differentially expressed genes and proteins indicated involvement in ER-related processes. The <em>washc5</em> knockout zebrafish model exhibits phenotypic similarities to human 3C syndrome, suggesting that mutations of this gene may play a pathogenic role in the syndrome. The mechanism of the <em>washc5</em> gene in 3C syndrome may be associated with disturbances in ER homeostasis, providing insights into potential gene therapy strategies.</div></div>","PeriodicalId":12499,"journal":{"name":"Gene","volume":"948 ","pages":"Article 149351"},"PeriodicalIF":2.6,"publicationDate":"2025-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143482741","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":"AhFAD3-A01 enhances α-linolenic acid content in Arabidopsis and peanut","authors":"Li Xiaona ,&nbsp;Xue Lulu ,&nbsp;Liu Han ,&nbsp;Qu Pengyu ,&nbsp;Zhao Huanhuan ,&nbsp;Luo Dandan ,&nbsp;Wang Xiaobo ,&nbsp;Huang Bingyan ,&nbsp;Zhang Maoning ,&nbsp;Li Chenyu ,&nbsp;Zhang Zhongxin ,&nbsp;Dong Wenzhao ,&nbsp;Shi Lei ,&nbsp;Zhang Xinyou","doi":"10.1016/j.gene.2025.149336","DOIUrl":"10.1016/j.gene.2025.149336","url":null,"abstract":"<div><div>Alpha-linolenic acid (ALA, C18:3) is an essential fatty acid integral to human growth and development. Despite its significance, the ALA content in peanut seeds-a major global oilseed crop-is notably low. This study employed bioinformatics analysis, tissue expression, and promoter function evaluations to investigate <em>AhFAD3</em>, which encodes the microsomal omega-3 fatty acid desaturase that is directly responsible for ALA accumulation through converting linoleic acid (LA) to ALA. We identified the active <em>AhFAD3</em> gene, <em>AhFAD3-A01</em>, with the functional protein encoded by <em>AhFAD3-A01</em> localized in the endoplasmic reticulum (ER) and found to be pivotal in ALA synthesis in seeds. The low expression of <em>AhFAD3</em> genes during the late stages of seed development, coupled with the specific activation by only <em>AhFAD3-A01</em> and <em>AhFAD3-B01</em> promoters in seeds, results in the low ALA levels in mature peanut seeds. To enhance ALA content, the constitutive promoter CaMV35S and the seed-specific promoter P<em>_AhWRI1</em> were utilized to overexpress <em>AhFAD3-A01</em> in Arabidopsis and peanut. While the expression level of <em>AhFAD3-A01</em> in P<em>_AhWRI1</em>::<em>AhFAD3-A01</em> transgenic Arabidopsis remained unchanged, it significantly increased under the CaMV35S::<em>AhFAD3-A01</em> configuration, leading to an over a 40 % increase in ALA content of in T₄ generation seeds, indicating that P<em>_AhWRI1</em> was unable to drive <em>AhFAD3</em> overexpression in Arabidopsis. Similarly, the overexpression of <em>AhFAD3-A01</em> using both promoters in peanuts resulted in enhanced expression and an increase in ALA content from 15.18 % to 30.65 % in CaMV35S::<em>AhFAD3-A01</em> T₁ generation seeds and from 11.23 % to 25.49 % in P<em>_AhWRI1</em>::<em>AhFAD3-A01</em> seeds. These results highlight the critical role of <em>AhFAD3-A01</em> ALA synthesis in peanut seeds and provide a solid foundation for developing peanut varieties with elevated ALA content.</div></div>","PeriodicalId":12499,"journal":{"name":"Gene","volume":"949 ","pages":"Article 149336"},"PeriodicalIF":2.6,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143476429","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":"Association of Zinc Finger-76 rs10947540 polymorphism and its gene expression in Egyptian patients with systemic lupus erythematosus","authors":"Ismail M. Ahmed ,&nbsp;Hanan Elimam ,&nbsp;Maha Emad Ibrahim ,&nbsp;Sarah Shabayek ,&nbsp;Dina M. Abo-Elmatty ,&nbsp;Noha M. Mesbah ,&nbsp;Asmaa R. Abdel-Hamed","doi":"10.1016/j.gene.2025.149343","DOIUrl":"10.1016/j.gene.2025.149343","url":null,"abstract":"<div><div>Systemic lupus erythematosus (SLE) is an autoimmune disorder characterized by the involvement of multiple organs and dysregulation of the immune system. Zinc finger (ZNF) proteins act as DNA-binding transcription factors, thereby regulating gene expression. Polymorphisms, specifically single-nucleotide polymorphisms (SNPs), within ZNF-coding genes, have been implicated in susceptibility to autoimmune diseases, including the ZNF-76 gene in SLE. This investigation aimed to elucidate the association between ZNF-76 gene expression, protein levels, and rs10947540 SNP in a cohort of Egyptian SLE patients. One hundred healthy controls and one hundred SLE patients were recruited. Quantitative real-time PCR was employed to identify ZNF-76 (C/T) rs10947540 genotypes and quantify serum ZNF-76 mRNA expression. Additionally, serum ZNF-76 protein levels were measured using ELISA. Our results revealed downregulation of ZNF-76 mRNA expression in approximately 75% of SLE patients, with a significant decrease (0.23-fold) in median expression compared to controls. Furthermore, SLE patients exhibited a higher prevalence of high-risk TT genotype and demonstrably lower serum ZNF-76 protein levels. In conclusion, this study suggests a strong association between decreased serum ZNF-76 mRNA expression and increased susceptibility to SLE in the studied Egyptian population. Specific SNP in the ZNF-76 gene may be potential markers for inherited predisposition to SLE in this population.</div></div>","PeriodicalId":12499,"journal":{"name":"Gene","volume":"948 ","pages":"Article 149343"},"PeriodicalIF":2.6,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143465535","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":"RNA research for drug discovery: Recent advances and critical insight","authors":"Patrick Maduabuchi Aja ,&nbsp;Peter Chinedu Agu ,&nbsp;Celestine Ogbu ,&nbsp;Esther Ugo Alum ,&nbsp;Ilemobayo Victor Fasogbon ,&nbsp;Angela Mumbua Musyoka ,&nbsp;Wisdom Ngwueche ,&nbsp;Chinedu Ogbonia Egwu ,&nbsp;Deusdedit Tusubira ,&nbsp;Kehinde Ross","doi":"10.1016/j.gene.2025.149342","DOIUrl":"10.1016/j.gene.2025.149342","url":null,"abstract":"<div><div>The field of RNA research has experienced significant changes and is now at the forefront of contemporary drug development. This narrative overview explores the scientific developments and historical turning points in RNA research, emphasising the field’s critical significance in the development of novel therapeutics. Important discoveries like antisense oligonucleotides (ASOs), mRNA therapies, and RNA interference (RNAi) have created novel treatment options that can be targeted, such as the ground-breaking mRNA vaccinations against COVID-19. Advances in high-throughput sequencing, single-cell RNA sequencing, and epitranscriptomics have further unravelled the complexity of RNA biology, shedding light on the intricacies of gene regulation and cellular diversity. The integration of computational tools and bioinformatics has propelled the identification of RNA-based biomarkers and the development of RNA therapeutics. Despite significant progress, challenges such as RNA stability, delivery, and off-target effects persist, necessitating continuous innovation and ethical considerations. This review provides a critical insight into the current state and prospects of RNA research, emphasising its transformative potential in drug discovery. By examining the interplay between technological advancements and therapeutic applications, we underscore the promising horizon for RNA-based interventions in treating a myriad of diseases, marking a new era in precision medicine.</div></div>","PeriodicalId":12499,"journal":{"name":"Gene","volume":"947 ","pages":"Article 149342"},"PeriodicalIF":2.6,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143463796","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":"Molecular Crosstalk by miR-449a and miR-34b in endometrial and ovarian cancer cells in vitro","authors":"Anuradha Sharma,&nbsp;Hanisha,&nbsp;Akanksha Rana,&nbsp;Indu Sharma","doi":"10.1016/j.gene.2025.149337","DOIUrl":"10.1016/j.gene.2025.149337","url":null,"abstract":"<div><h3>Aims</h3><div>Endometrial and ovarian cancers, the sixth and eighth most prevalent cancers in women globally, account for nearly 8% of all new female cancer cases annually. MicroRNAs (miRNAs) have emerged as a promising field in cancer treatment, offering new avenues for targeted therapies and diagnostic tools. Recent miRNA-based cancer research has uncovered various miRNAs commonly dysregulated in cancer and which possess tumor-suppressive functions. These miRNAs influence genes crucial for cellular differentiation, proliferation, apoptosis, and metabolism.</div></div><div><h3>Main methods</h3><div>In the present study, the researchers investigated the effect of dysregulation of two such miRNAs, miR-449a and miR-34b, on the oncogenes involved in the progression of endometrial and ovarian cancer using the respective RL95-2 and SKOV3 cell lines. The transcriptional gene expression analysis was done by Real-Time Quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR).</div></div><div><h3>Key findings</h3><div>It was found that the overexpression of miR-449a and miR-34b downregulated <em>HIF-1α, VEGF, c-Myc</em>, <em>COX-2</em>, and <em>TNF-α</em> while upregulating <em>TP53</em> in both cancer types. Conversely, inhibiting these miRNAs increased the levels of <em>HIF-1α, VEGF, c-Myc, COX-2</em>, and <em>TNF-α</em>, and decreased <em>TP53</em>. However, co-transfection with both mimic and inhibitor had varying effects.</div></div><div><h3>Significance</h3><div>The study demonstrated that these miRNAs could influence critical processes such as angiogenesis, proliferation, inflammation, tumorigenesis, and apoptosis in cancer cells, highlighting their potential as therapeutic targets. However, the varied effects observed with the co-transfection of mimics and inhibitors suggest a complex interplay that requires further investigation.</div></div>","PeriodicalId":12499,"journal":{"name":"Gene","volume":"947 ","pages":"Article 149337"},"PeriodicalIF":2.6,"publicationDate":"2025-02-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143438276","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":"Crosstalk between signaling pathways (Rho/ROCK, TGF-β and Wnt/β-Catenin Pathways/ PI3K-AKT-mTOR) in Cataract: A Mechanistic Exploration and therapeutic strategy","authors":"Meraj Khan,&nbsp;Lokesh Verma","doi":"10.1016/j.gene.2025.149338","DOIUrl":"10.1016/j.gene.2025.149338","url":null,"abstract":"<div><div>Cataract are a leading cause of visual impairment that is characterized by clouding or lens opacification of the healthy clear lens of the eye or its capsule. It can be classified based on their etiology and clinical presentation such as congenital, age-related, and secondary cataracts. Clinically, it may be further classified as a cortical or nuclear cataract. Cortical cataracts are responsible for opacification of the lens cortex, while nuclear cataracts cause age-related degeneration of the lens nucleus. This review aims to explore the molecular mechanism associated with various signaling pathways underlying cataract formation. Additionally, explore the potential therapeutic strategies for the management of cataracts. A comprehensive literature search was performed utilizing different keywords such as cataract, pathogenesis, signaling pathways, therapeutic approaches, RNA therapeutics, and surgery. Electronic databases such as PubMed, Google Scholar, Springer Link, and Web of Science were used for the literature search. The cataract formation is responsible for protein aggregation, primarily of γ-crystallin, and causes disruptions in signaling pathways. Key pathways include Rho/ROCK, TGF-β, Wnt/β-catenin, NF-κB, and PI3K-AKT-mTOR. Signaling pathways governing lens epithelial cell differentiation and epithelial-to-mesenchymal transition (EMT) are essential for maintaining lens transparency. Disruptions in these pathways, often caused by genetic mutations in genes like MIP, TDRD7, PAX6, FOXE3, HSF4, MAF, and PITX3 lead to cataract formation. While surgical intervention remains the primary treatment, pharmacological therapies and emerging RNA-based strategies offer promising strategies for the prevention and management of cataracts. A deeper understanding of the underlying molecular mechanisms is essential to develop innovative therapeutic strategies and improve the quality of life for individuals affected by cataracts.</div></div>","PeriodicalId":12499,"journal":{"name":"Gene","volume":"947 ","pages":"Article 149338"},"PeriodicalIF":2.6,"publicationDate":"2025-02-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143448770","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":"GANAB c.1118C > T is a novel variant in patients with polycystic liver disease / polycystic kidney disease","authors":"Xiuling Zhu ,&nbsp;Ying Liang ,&nbsp;Xiaoling Zhou ,&nbsp;Zhiqiang Zhang,&nbsp;Yuzhen Liu,&nbsp;Zhongyuan Cui,&nbsp;Zhixian Wu,&nbsp;Dongliang Li","doi":"10.1016/j.gene.2025.149339","DOIUrl":"10.1016/j.gene.2025.149339","url":null,"abstract":"<div><h3>Background</h3><div>Many patients suffer from genetically unsolved polycystic liver disease (PLD). Our aim was to explore mutated genes associated with the pathogenesis of PLD.</div></div><div><h3>Methods</h3><div>First, a family investigation was conducted on probands with a definite diagnosis of PLD and the pedigree was confirmed. Data on the clinical symptoms and biochemical and imaging indicators of family members were collected. Mutation analysis was performed using whole exome sequencing (WES), and Sanger sequencing was used for mutation verification.</div></div><div><h3>Results</h3><div>Nine of the 33 patients from five generations of the proband and their families were diagnosed with PLD/polycystic kidney disease (PKD). Imaging examination of all patients confirmed numerous hepatic and renal cysts. Patients clinically presented with different degrees of abdominal distension and impaired renal function. WES revealed a missense mutation in <em>GANAB</em> c.1118C &gt; T (p. Thr373Ile) in the proband and her affected son, which resulted in a change from hydrophilic threonine to hydrophobic isoleucine at amino acid 373. The mutation was verified using Sanger sequencing.</div></div><div><h3>Conclusion</h3><div>This study identified a novel heterozygous mutation, c.1118C &gt; T (p. Thr373Ile) in <em>GANAB</em> in patients with PLD/PKD. This mutation has not been reported to date and enriches the phenotype and genotype spectrum of <em>GANAB</em>-related diseases.</div></div>","PeriodicalId":12499,"journal":{"name":"Gene","volume":"946 ","pages":"Article 149339"},"PeriodicalIF":2.6,"publicationDate":"2025-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143428747","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":"Leishmania donovani homoserine dehydrogenase: Biochemical and structural characterization of a novel parasite specific enzyme of aspartate pathway","authors":"Sandra Lalchhuanawmi ,&nbsp;Chandan Kumar Malik ,&nbsp;Rajender Kumar ,&nbsp;Swati Verma ,&nbsp;Prabha Garg ,&nbsp;Sushma Singh","doi":"10.1016/j.gene.2025.149335","DOIUrl":"10.1016/j.gene.2025.149335","url":null,"abstract":"<div><div>Visceral leishmaniasis is a neglected tropical disease. Drug resistance and toxicity are the critical issues with the currently available antileishmanial drugs. Therefore, research efforts are underway to identify and validate new drug targets specific to Leishmania parasite. The enzyme homoserine dehydrogenase (HSD) functions in the third step of aspartate pathway. The present study focuses on the biophysical and biochemical characterization of HSD enzyme from <em>Leishmania donovani</em> (<em>Ld</em>HSD) which is unique to the parasite with no homologous enzyme in the host. <em>Ld</em>HSD gene was cloned in pET28c(+) vector and transformed in <em>E. coli</em> BL21 (DE3) strain. <em>Ld</em>HSD recombinant enzyme of molecular weight 46.6 kDa with 6X-His tag at the C-terminal end was expressed, purified by nickel affinity chromatography and confirmed by western blot analysis using anti-His antibody. Effect of pH, temperature, salts, metal ions and amino acids on the recombinant enzyme were evaluated. Kinetic parameters of <em>Ld</em>HSD were evaluated for substrates L-homoserine and NADP⁺. Biophysical analysis revealed that the enzyme is rich in β-sheets. Thermal denaturation study revealed that the protein is stable up to 45 °C. Furthermore, comprehensive comparative sequence analysis and structural modeling revealed the structural and functionally important residues, which are involved in the catalytic mechanisms. The putative binding mode of the natural substrate L-homoserine into the active site of <em>Ld</em>HSD was also elucidated. These findings provide a foundation for the development of selective, target-based inhibitors against the HSD enzyme of the parasite.</div></div>","PeriodicalId":12499,"journal":{"name":"Gene","volume":"947 ","pages":"Article 149335"},"PeriodicalIF":2.6,"publicationDate":"2025-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143440540","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}