WoS İndeksli Yayınlar Koleksiyonu / WoS Indexed Publications Collection
Permanent URI for this collectionhttps://hdl.handle.net/11147/7150
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Article Citation - WoS: 4Citation - Scopus: 3Expression Patterns of M6a Rna Methylation Regulators Under Apoptotic Conditions in Various Human Cancer Cell Lines(TUBITAK, 2024) Alasar, Azime Akçaöz; Sağlam, Buket; Vatansever, İpek Erdoğan; Akgül, BünyaminBackground/aim: Cancer is a complex disease that involves both genetic and epigenetic factors. While emerging evidence clearly suggests that changes in epitranscriptomics play a crucial role in cancer pathogenesis, a comprehensive understanding of the writers, erasers, and readers of epitranscriptomic processes, particularly under apoptotic conditions remains lacking. The aim of this study was to uncover the changes in the expression of m6A RNA modifiers under apoptotic conditions across various cancer cell lines. Materials and methods: Initially, we quantified the abundance of m6A RNA modifiers in cervical (HeLa and ME180), breast (MCF7 and MDA-MB-231), lung (A549 and H1299), and colon (Caco-2 and HCT116) cancer cell lines using qPCR. Subsequently, we induced apoptosis using cisplatin and tumor necrosis factor-alpha (TNF-α) to activate intrinsic and extrinsic pathways, respectively, and assessed apoptosis rates via flow cytometry. Further, we examined the transcript abundance of m6A RNA modifiers under apoptotic conditions in cervical, breast, and lung cancer cell lines using qPCR. Results: Overall, treatment with cisplatin increased the abundance of m 6A modifiers, whereas TNF-α treatment decreased their expression in cervical, breast, and lung cancer cell lines. Specifically, cisplatin-induced apoptosis, but not TNF-α-mediated apoptosis, resulted in decreased abundance of METTL14 and FTO transcripts. Additionally, cisplatin treatment drastically reduced the abundance of IGF2BP2 and IGF2BP3 readers. Conclusion: These results suggest that the differential response of cancer cells to apoptotic inducers may be partially attributed to the expression of m6A RNA modifiers.Article Citation - WoS: 28Citation - Scopus: 30Bone Marrow Stem Cells Adapt To Low-Magnitude Vibrations by Altering Their Cytoskeleton During Quiescence and Osteogenesis(TUBITAK, 2015) Demiray, Levent; Özçivici, EnginApplication of mechanical vibrations is anabolic to bone tissue, not only by guiding mature bone cells to increased formation, but also by increasing the osteogenic commitment of progenitor cells. However, the sensitivity and adaptive response of bone marrow stem cells to this loading regimen has not yet been identified. In this study, we subjected mouse bone marrow stem cell line D1-ORL-UVA to daily mechanical vibrations (0.15 g, 90 Hz, 15 min/day) for 7 days, both during quiescence and osteogenic commitment, to identify corresponding ultrastructural adaptations on cellular and molecular levels. During quiescence, mechanical vibrations significantly increased total actin content and actin fiber thickness, as measured by phalloidin staining and fluorescent microscopy. Cellular height also increased, as measured by atomic force microscopy, along with the expression of focal adhesion kinase (PTK2) mRNA levels. During osteogenesis, mechanical vibrations increased the total actin content, actin fiber thickness, and cytoplasmic membrane roughness, with significant increase in Runx2 mRNA levels. These results show that bone marrow stem cells demonstrate similar cytoskeletal adaptations to low-magnitude high-frequency mechanical loads both during quiescence and osteogenesis, potentially becoming more sensitive to additional loads by increased structural stiffness.Article Citation - WoS: 6Citation - Scopus: 8Deep Sequencing Reveals Two Jurkat Subpopulations With Distinct Mirna Profiles During Camptothecin-Induced Apoptosis(TUBITAK, 2018) Erdoğan, İpek; Coşacak, Mehmet İlyas; Nalbant, Ayten; Akgül, BünyaminMicroRNAs (miRNAs) are small noncoding RNAs of about 19-25 nt that regulate gene expression posttranscriptionally under various cellular conditions, including apoptosis. The miRNAs involved in modulation of apoptotic events in T cells are partially known. However, heterogeneity associated with cell lines makes it difficult to interpret gene expression signatures, especially in cancer-related cell lines. Treatment of the Jurkat T-cell leukemia cell line with the universal apoptotic drug, camptothecin, resulted in identification of two Jurkat subpopulations: one that is sensitive to camptothecin and another that is rather intrinsically resistant. We sorted apoptotic Jurkat cells from nonapoptotic ones prior to profiling miRNAs through deep sequencing. Our data showed that a total of 184 miRNAs were dysregulated. Interestingly, the apoptotic and nonapoptotic subpopulations exhibited distinct miRNA expression profiles. In particular, 6 miRNAs were inversely expressed in these two subpopulations. The pyrosequencing results were validated by real-time qPCR. Altogether, these results suggest that miRNAs modulate apoptotic events in T cells and that cellular heterogeneity requires careful interpretation of miRNA expression profiles obtained from drug-treated cell lines.Article Citation - WoS: 1Citation - Scopus: 1Comparative Proteomic Analysis of Bacillus Thuringiensis Wild-Type and Two Mutant Strains Disturbed in Polyphosphate Homeostasis(TUBITAK, 2018) Yeşilırmak, Filiz; Doruk, Tuğrul; Yılmaz, Şerif; Tunca Gedik, SedefPolyphosphate polymer (polyP) plays a very important role in every living cell. Synthesis of this linear polymer of phosphate (Pi) residues is catalyzed by the polyphosphate kinase (PPK) enzyme. It was shown that high levels of intracellular polyphosphate stimulated endotoxin production by Bacillus thuringiensis subsp. israelensis (Bti). In this study, proteomic analysis of the wild-type and two mutant strains, overexpressing the ppk gene (Bti pHTppk) and without the ppk gene (Bti ∆ppk), were used to clarify the relation between polyP and endotoxin production. Intracellular proteins were separated by two-dimensional gel electrophoresis; 41 spots of interest (proteins differentially expressed) were obtained and 35 of them were identified by mass spectrometry. Analysis of the protein profiles showed that there is a general decrease in the expression levels of proteins related with energy metabolism, amino acid metabolism, and purine biosynthesis in both Bti pHTppk and Bti ∆ppk. Gluconeogenesis and fatty acid metabolism were also slowed down in both strains, whereas expression of stress response proteins increased compared to the wild-type. These results suggested that changes in polyP concentration cause a general stress condition inside the cell, which in turn stimulates secondary metabolite synthesisArticle Citation - WoS: 3Citation - Scopus: 3Cloning, Expression, and Activity Analysis of Human Cathepsin C in the Yeast Pichia Pastoris(TUBITAK, 2017) Dağlıoğlu, CenkThe yeast Pichia pastoris expression system was investigated for the production of human cathepsin C (CatC) recombinant protein. The full-length CatC cDNA, corresponding to amino acids 12-475, was synthesized from interleukin-2 (IL-2) stimulated human peripheral blood mononuclear cells and subcloned in the pGEM-T cloning vector. After confirming the DNA sequence of the insert, the gene was cloned into the pPICZαA expression vector under the control of the methanol-inducible alcohol oxidase (AOX1) promoter and transformed to P. pastoris X-33 cells. The expressed protein was secreted into the culture medium through the α-factor mating signal sequence of the expression vector. Analysis of the culture supernatant revealed that the recombinant human CatC was secreted as a 58-kDa molecule, indicating that human CatC was accumulated in the culture supernatant as proform composed of the residual propart, the activation peptide, and the heavy and light chains. Extracellular recombinant proCatC was further activated by cysteine endoprotease papain in vitro and its activity was confirmed by assays using a synthetic substrate.Article Citation - WoS: 13Citation - Scopus: 14Evaluation of Orange Peel, an Industrial Waste, for the Production of Aspergillus Sojae Polygalacturonase Considering Both Morphology and Rheology Effects(TUBITAK, 2014) Gögüş, Nihan; Hakgüder Taze, Bengi; Demir, Hande; Tarı, Canan; Ünlütürk, Sevcan; Lahore, Marcelo FernandezOrange peel is an agroindustrial waste rich in pectin and known to be an inducer for pectinase production. The use of this low-cost substrate for the production of an industrially important enzyme, polygalacturonase (PG), can be an alternative way to turn this waste into a value-added product, contributing to the reduction of environmental waste disposal problems. Enzyme productions by fungal microorganisms are affected by environmental and nutritional factors, demanding the determination of optimum conditions for maximum enzyme production with the desired fungal morphology and broth rheology. Therefore, complex and additional carbon sources were optimized with respect to PG production by Aspergillus sojae using statistical approaches. Effect of pH, another significant parameter affecting the rheology and morphology of the strain, was investigated in the serial bioreactor system using the optimized medium composition. Highest PG enzyme yield and productivity together with the maximum PG enzyme production (93.48 U/mL) were obtained under uncontrolled pH conditions. Under these conditions, morphologically, pellet sizes exhibited a normal distribution ranging between 0.5-1.0 mm and 1.0-1.5 mm, and rheological measurements revealed that fermentation broths showed non-Newtonian flow. The low pH trend observed during the course of the fermentation was another important positive outcome for industrial fermentations, prone to contamination problems.Article Citation - WoS: 5Citation - Scopus: 5Invadopodia: Proteolytic Feet of Cancer Cells(TUBITAK, 2014) Batı, Gizem; Pesen Okvur, DevrimThe leading cause of death in cancer patients is metastasis. Invasion is an integral part of metastasis and is carried out by proteolytic structures called invadopodia at the cellular level. In this introductory review, we start by evaluating the definition of invadopodia. While presenting the upstream signaling events involved, we integrate current models on invadopodia. In addition, we discuss the significance of invadopodia in 2D and 3D and in vivo. We finally point out technical challenges and conclude with open questions in the field. © TÜBİTAK.Article Citation - WoS: 28Citation - Scopus: 32Optimization of the Process Parameters for the Utilization of Orange Peel To Produce Polygalacturonase by Solid-State Fermentation From an Aspergillus Sojae Mutant Strain(TUBITAK, 2012) Demir, Hande; Göğüş, Nihan; Tarı, Canan; Heerd, Doreen; Lahore, Marcelo FernandezThe effect of orange peel concentration, HCl concentration, incubation time and temperature, and inoculum size on the spore count and activity of polygalacturonase (PG) enzyme produced from Aspergillus sojae M3 by solidstate fermentation was screened using 2k factorial design. Orange peel and HCl concentrations and incubation time were significant factors affecting the responses. Optimum conditions favoring both PG and spore production from Aspergillus sojae M3 were determined as 2% orange peel and 50 mM HCl concentrations at 22 °C and 4.3 days of incubation. An overlay plot was constructed for use as a practical chart for production of high enzyme activity (>35.0 U/g substrate) and spore count (9.0 × 108 to 2.0 × 109 spore/mL) by superimposing the contours of PG activity and spore count responses. The accuracy and reliability of the constructed models on the responses was validated with the maximum calculated error rate between the predicted and actual activities at 14.1% and 22.4%, respectively. © TÜBİTAK.Article Citation - WoS: 27Citation - Scopus: 36Purification and Biochemical Characterization of an Extracellular Lipase From Psychrotolerant Pseudomonas Fluorescens Ke38(TUBITAK, 2013) Adan Gökbulut, Aysun; Arslanoğlu, AlperAn extracellular lipase producing bacterium was isolated from a soil sample, and identified as a strain of Pseudomonas fluorescens by 16S rRNA gene sequencing. It was named Pseudomonas fluorescens KE38. KE38 showed psychrotolerant properties with an optimum growth temperature of 25 °C. The lipase enzyme secreted by KE38 was purified 41.13-fold with an overall yield of 54.99%, and a specific activity of 337.3 U/mg. The molecular mass of purified lipase was estimated to be approximately 43 kDa by SDS-PAGE. Although the lipase was active at a temperature range of 15-65 °C, it exhibited maximum activity at 45 °C, at pH 8.0. The enzyme exhibited high stability retaining 100% and 70% of its activity after an incubation period of 45 and 100 min at 45 °C and pH 8.0 respectively. It also showed a broad substrate specificity acting on p-nitrophenyl esters with C8-C18 acyl groups as substrates and was activated by Ca2+ and Ni2+ at 1 mM. While the enzyme retained its activity levels in the presence of a variety of organic solvents, DMSO and dimethylformamide enhanced this. High stability, broad substrate specificity and activity at cold temperatures in the presence of organic solvents, and metal ions make the extracellular lipase of KE38 a candidate for industrial applications.Article Citation - WoS: 25Citation - Scopus: 25Application of Est-Ssrs To Examine Genetic Diversity in Eggplant and Its Close Relatives(TUBITAK, 2011) Tümbilen, Yeliz; Frary, Anne; Daunay, Marie Christine; Doğanlar, SamiWithin the genus Solanum, the term 'eggplant' encompasses several cultivated species that are used for food and, to a lesser extent, for medicine. Th e use of one common name to describe more than one species and the existence of many related wild species have led to taxonomic confusion which, in turn, have complicated analyses of evolutionary relationships and genetic diversity within this groups of species. A further challenge for eggplant research is that, despite the fact that the use of molecular markers for phylogenetic studies is well-established, very few studies have described the development of new markers for eggplant. In our work, genic microsatellite (SSR) markers were identified from an expressed sequence tag library of S. melongena and used for analysis of 47 accessions of eggplant and closely related species. Th e markers had very good polymorphism in the 18 species tested including 8 S. melongena accessions. Moreover, genetic analysis performed with these markers showed concordance with previous research and knowledge of eggplant domestication. Th ese markers are expected to be a valuable resource for studies of genetic relationships, fingerprinting, and gene mapping in eggplant.