Lung cancer is one of the cancer types with the highest mortality worldwide. The most frequently mutated genes known to be clinically important in lung cancers are EGFR, BRAF, and KRAS genes. Therefore, the therapeutic agents developed are directed against variants that cause over-activation of the EGFR–KRAS–BRAF–BRAF–MEK/ERK signalling pathway. However, different responses of patients to Tyrosine Kinase Inhibitors (TKIs) suggest that new prognostic biomarkers should be defined and epigenetic mechanisms may be related to this situation. Methods: In this study, sequence analyses of AGO2, DICER, and DROSHA genes involved in miRNA biogenesis and EGFR, KRAS, and BRAF genes were performed in 35 patients with sporadic lung cancer. Results: We found variations in genes involved in miRNA biogenesis that have not been previously reported in the literature. In addition, we found 4 different variants in the EGFR gene that have been described in the literature. In addition, a statistically significant association was found between the presence of mutations in at least one of the genes involved in miRNA biogenesis and metastasis (p:0.02). Conclusions: In conclusion, genomic dysregulation of key miRNA biogenesis genes may be one of the possible reasons for the differential response of patients to therapeutic agents and the development of metastasis in EGFR wild type tumours.
{"title":"Variant Analysis of miRNA Regulatory Genes in 35 Sporadic Lung Carcinoma Tumors","authors":"Özkan Bağcı, Ebru Marzioğlu Özdemir, Batuhan Şanlıtürk","doi":"10.1134/S1607672924600052","DOIUrl":"10.1134/S1607672924600052","url":null,"abstract":"<p>Lung cancer is one of the cancer types with the highest mortality worldwide. The most frequently mutated genes known to be clinically important in lung cancers are <i>EGFR</i>, <i>BRAF</i>, and <i>KRAS</i> genes. Therefore, the therapeutic agents developed are directed against variants that cause over-activation of the EGFR–KRAS–BRAF–BRAF–MEK/ERK signalling pathway. However, different responses of patients to Tyrosine Kinase Inhibitors (TKIs) suggest that new prognostic biomarkers should be defined and epigenetic mechanisms may be related to this situation. Methods: In this study, sequence analyses of <i>AGO2</i>, <i>DICER</i>, and <i>DROSHA</i> genes involved in miRNA biogenesis and <i>EGFR</i>, <i>KRAS</i>, and <i>BRAF</i> genes were performed in 35 patients with sporadic lung cancer. Results: We found variations in genes involved in miRNA biogenesis that have not been previously reported in the literature. In addition, we found 4 different variants in the <i>EGFR</i> gene that have been described in the literature. In addition, a statistically significant association was found between the presence of mutations in at least one of the genes involved in miRNA biogenesis and metastasis (p:0.02). Conclusions: In conclusion, genomic dysregulation of key miRNA biogenesis genes may be one of the possible reasons for the differential response of patients to therapeutic agents and the development of metastasis in EGFR wild type tumours.</p>","PeriodicalId":529,"journal":{"name":"Doklady Biochemistry and Biophysics","volume":"513 1 supplement","pages":"S1 - S7"},"PeriodicalIF":0.8,"publicationDate":"2024-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140108706","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-03-12DOI: 10.1134/S1607672924700765
V. E. Balakin, T. A. Belyakova, O. M. Rozanova, E. N. Smirnova, N. S. Strelnikova, E. A. Kuznetsova
The effect of carbon ions (12C) with the energy of 400 MeV/nucleon on the dynamics of induction and growth rate of solid tumors in mice under irradiation of Ehrlich ascites carcinoma cells (EAC) ex vivo at doses of 5–30 Gy relative to the action of equally effective doses of X-ray radiation was studied. The dynamics of tumor induction under the action of 12C and X-rays had a similar character and depended on the dose during 3 months of observation. The value of the latent period, both when irradiating cells with 12C and X-ray, increased with increasing dose, and the interval for tumor induction decreased. The rate of tumor growth after ex vivo irradiation of EAC cells was independent of either dose or type of radiation. The dose at which EAC tumors are not induced within 90 days was 30 Gy for carbon ions and 60 Gy for X-rays. The value of the relative biological effectiveness of carbon ions, calculated from an equally effective dose of 50% probability of tumors, was 2.59.
研究了能量为 400 MeV/nucleon的碳离子(12C)对小鼠体内外照射艾氏腹水癌细胞(EAC)时实体瘤诱导和生长速度的动态影响,照射剂量为 5-30 Gy,相对于同样有效剂量的 X 射线辐射作用。在 3 个月的观察期间,12C 和 X 射线作用下肿瘤诱导的动态具有相似的特征,并取决于剂量。用 12C 和 X 射线照射细胞时,潜伏期的值都随着剂量的增加而增加,肿瘤诱导的间隔时间则缩短。EAC细胞体外照射后的肿瘤生长率与剂量或辐射类型无关。90 天内不诱发 EAC 肿瘤的剂量为碳离子 30 Gy 和 X 射线 60 Gy。根据肿瘤概率为 50%的同等有效剂量计算,碳离子的相对生物有效性值为 2.59。
{"title":"Anti-tumor Effect of High Doses of Carbon Ions and X-Rays during Irradiation of Ehrlich Ascites Carcinoma Cells Ex Vivo","authors":"V. E. Balakin, T. A. Belyakova, O. M. Rozanova, E. N. Smirnova, N. S. Strelnikova, E. A. Kuznetsova","doi":"10.1134/S1607672924700765","DOIUrl":"10.1134/S1607672924700765","url":null,"abstract":"<p>The effect of carbon ions (<sup>12</sup>C) with the energy of 400 MeV/nucleon on the dynamics of induction and growth rate of solid tumors in mice under irradiation of Ehrlich ascites carcinoma cells (EAC) ex vivo at doses of 5–30 Gy relative to the action of equally effective doses of X-ray radiation was studied. The dynamics of tumor induction under the action of <sup>12</sup>C and X-rays had a similar character and depended on the dose during 3 months of observation. The value of the latent period, both when irradiating cells with <sup>12</sup>C and X-ray, increased with increasing dose, and the interval for tumor induction decreased. The rate of tumor growth after ex vivo irradiation of EAC cells was independent of either dose or type of radiation. The dose at which EAC tumors are not induced within 90 days was 30 Gy for carbon ions and 60 Gy for X-rays. The value of the relative biological effectiveness of carbon ions, calculated from an equally effective dose of 50% probability of tumors, was 2.59.</p>","PeriodicalId":529,"journal":{"name":"Doklady Biochemistry and Biophysics","volume":"513 1 supplement","pages":"S30 - S35"},"PeriodicalIF":0.8,"publicationDate":"2024-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140108655","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-03-12DOI: 10.1134/S1607672924700753
L. A. Ovchinnikova, S. S. Dzhelad, T. O. Simaniv, M. N. Zakharova, A. G. Gabibov, Y. A. Lomakin
Multiple sclerosis (MS) is an autoimmune neurodegenerative disease leading to inevitable disability and primarily affecting the young and middle-aged population. Recent studies have shown a direct correlation between the risk of MS development and Epstein–Barr virus (EBV) infection. Analysis of the titer of EBV-specific antibodies among patients with MS and healthy donors among Russian population confirmed that MS is characterized by an increased level of serum IgG binding EBNA-1 (EBV nuclear antigen 1). The number of patients with elevated levels of EBNA-1-specific antibodies does not differ statistically significantly between two groups with diametrically opposite courses of MS: benign MS or highly active MS. It can be assumed that the primary link between EBV and the development of MS is restricted to the initiation of the disease and does not impact its severity.
{"title":"The Level of Anti-Viral Antigen-Specific Antibodies to EBNA-1 in the Serum of MS Patients Does not Depend on the Severity of the Disease","authors":"L. A. Ovchinnikova, S. S. Dzhelad, T. O. Simaniv, M. N. Zakharova, A. G. Gabibov, Y. A. Lomakin","doi":"10.1134/S1607672924700753","DOIUrl":"10.1134/S1607672924700753","url":null,"abstract":"<p>Multiple sclerosis (MS) is an autoimmune neurodegenerative disease leading to inevitable disability and primarily affecting the young and middle-aged population. Recent studies have shown a direct correlation between the risk of MS development and Epstein–Barr virus (EBV) infection. Analysis of the titer of EBV-specific antibodies among patients with MS and healthy donors among Russian population confirmed that MS is characterized by an increased level of serum IgG binding EBNA-1 (EBV nuclear antigen 1). The number of patients with elevated levels of EBNA-1-specific antibodies does not differ statistically significantly between two groups with diametrically opposite courses of MS: benign MS or highly active MS. It can be assumed that the primary link between EBV and the development of MS is restricted to the initiation of the disease and does not impact its severity.</p>","PeriodicalId":529,"journal":{"name":"Doklady Biochemistry and Biophysics","volume":"515 1","pages":"48 - 51"},"PeriodicalIF":0.8,"publicationDate":"2024-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11021220/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140108705","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-03-12DOI: 10.1134/S1607672924700741
Y. V. Pekina, V. A. Babosha, P. G. Georgiev, A. A. Fedotova
In Drosophila, a large group of actively transcribed genes is located in pericentromeric heterochromatin. It is assumed that heterochromatic proteins recruit transcription factors to gene promoters. Two proteins, Ouib and Nom, were previously shown to bind to the promoters of the heterochromatic genes nvd and spok. Interestingly, Ouib and Nom are paralogs of the M1BP protein, which binds to the promoters of euchromatic genes. We have shown that, like M1BP, the Quib and Nom proteins bind to CP190, which is involved in the recruitment of transcription complexes to promoters. Unlike heterochromatic proteins, Ouib and Nom do not interact with the major heterochromatic protein HP1a and bind to euchromatic promoters on polytene chromosomes from the larval salivary glands. The results suggest a new mechanism for the recruitment of transcription factors into the heterochromatic compartment of the nucleus.
在果蝇中,一大批活跃的转录基因位于中心粒周围的异染色质中。据推测,异染色质蛋白将转录因子招募到基因启动子上。之前有研究表明,Ouib 和 Nom 这两种蛋白能与异染色质基因 nvd 和 spok 的启动子结合。有趣的是,Ouib 和 Nom 是 M1BP 蛋白的旁系亲属,而 M1BP 蛋白能与外染色质基因的启动子结合。我们已经证明,与 M1BP 蛋白一样,Quib 和 Nom 蛋白也与 CP190 结合,而 CP190 参与转录复合体与启动子的招募。与异染色质蛋白不同的是,Ouib 和 Nom 不与主要的异染色质蛋白 HP1a 相互作用,而是与幼虫唾液腺多体染色体上的非染色质启动子结合。这些结果表明,转录因子被招募到细胞核的异染色区是一种新的机制。
{"title":"Study of the Association of Ouib and Nom with Heterochromatin in Drosophila melanogaster","authors":"Y. V. Pekina, V. A. Babosha, P. G. Georgiev, A. A. Fedotova","doi":"10.1134/S1607672924700741","DOIUrl":"10.1134/S1607672924700741","url":null,"abstract":"<p>In <i>Drosophila</i>, a large group of actively transcribed genes is located in pericentromeric heterochromatin. It is assumed that heterochromatic proteins recruit transcription factors to gene promoters. Two proteins, Ouib and Nom, were previously shown to bind to the promoters of the heterochromatic genes <i>nvd</i> and <i>spok</i>. Interestingly, Ouib and Nom are paralogs of the M1BP protein, which binds to the promoters of euchromatic genes. We have shown that, like M1BP, the Quib and Nom proteins bind to CP190, which is involved in the recruitment of transcription complexes to promoters. Unlike heterochromatic proteins, Ouib and Nom do not interact with the major heterochromatic protein HP1a and bind to euchromatic promoters on polytene chromosomes from the larval salivary glands. The results suggest a new mechanism for the recruitment of transcription factors into the heterochromatic compartment of the nucleus.</p>","PeriodicalId":529,"journal":{"name":"Doklady Biochemistry and Biophysics","volume":"513 1 supplement","pages":"S26 - S29"},"PeriodicalIF":0.8,"publicationDate":"2024-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140108658","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-03-12DOI: 10.1134/S1607672924700777
Y. E. Zharkov, S. T. Zhorzholiani, A. A. Sergeev, A. V. Agafonov, A. Y. Gorodkov, L. A. Bockeria
Study of swirling flows in channels corresponding to the static approximation of flow channels of the heart and major vessels with a longitudinal–radial profile zR2 = const and a concave streamlined surface at the beginning of the longitudinal coordinate has been carried out. A comparative analysis of the flow structure in channel configurations zRN = const, where N = –1, 1, 2, 3, in the absence and presence of a concave surface was carried out. The numerical modeling was compared with the results of hydrodynamic experiments on the flow characteristics and the shape of the flow lines. The numerical model was used to determine the velocity structure, viscous friction losses, and shear stresses. Numerical modeling of steady-state flows for channels without a concave surface showed that in the channel zR2 = const there is a stable vortex flow structure with the lowest viscous friction losses. The presence of a concave surface of sufficient size significantly reduces viscous friction losses and shear stresses in both the steady state and pulsed modes.
{"title":"Experimental and Model Study of a Swirling Fluid Flow in a Converging Channel As a Simulation of Blood Flow in the Heart and Aorta","authors":"Y. E. Zharkov, S. T. Zhorzholiani, A. A. Sergeev, A. V. Agafonov, A. Y. Gorodkov, L. A. Bockeria","doi":"10.1134/S1607672924700777","DOIUrl":"10.1134/S1607672924700777","url":null,"abstract":"<p>Study of swirling flows in channels corresponding to the static approximation of flow channels of the heart and major vessels with a longitudinal–radial profile <i>zR</i><sup>2</sup> = const and a concave streamlined surface at the beginning of the longitudinal coordinate has been carried out. A comparative analysis of the flow structure in channel configurations <i>zR</i><sup><i>N</i></sup> = const, where <i>N</i> = –1, 1, 2, 3, in the absence and presence of a concave surface was carried out. The numerical modeling was compared with the results of hydrodynamic experiments on the flow characteristics and the shape of the flow lines. The numerical model was used to determine the velocity structure, viscous friction losses, and shear stresses. Numerical modeling of steady-state flows for channels without a concave surface showed that in the channel <i>zR</i><sup>2</sup> = const there is a stable vortex flow structure with the lowest viscous friction losses. The presence of a concave surface of sufficient size significantly reduces viscous friction losses and shear stresses in both the steady state and pulsed modes.</p>","PeriodicalId":529,"journal":{"name":"Doklady Biochemistry and Biophysics","volume":"513 1 supplement","pages":"S36 - S52"},"PeriodicalIF":0.8,"publicationDate":"2024-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140108656","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-03-12DOI: 10.1134/S1607672923700722
N. M. Shepelev, A. O. Kurochkina, O. A. Dontsova, M. P. Rubtsova
High-throughput ribosome profiling demonstrates the translation of thousands of small open reading frames located in the 5′ untranslated regions of messenger RNAs (upstream ORFs). Upstream ORF can both perform a regulatory function by influencing the translation of the downstream main ORF and encode a small functional protein or microprotein. In this work, we showed that the 5′ untranslated region of the PRPF19 mRNA encodes an upstream ORF that is translated in human cells. Inactivation of this upstream ORF reduces the viability of human cells.
{"title":"PRPF19 mRNA Encodes a Small Open Reading Frame That Is Important for Viability of Human Cells","authors":"N. M. Shepelev, A. O. Kurochkina, O. A. Dontsova, M. P. Rubtsova","doi":"10.1134/S1607672923700722","DOIUrl":"10.1134/S1607672923700722","url":null,"abstract":"<p>High-throughput ribosome profiling demonstrates the translation of thousands of small open reading frames located in the 5′ untranslated regions of messenger RNAs (upstream ORFs). Upstream ORF can both perform a regulatory function by influencing the translation of the downstream main ORF and encode a small functional protein or microprotein. In this work, we showed that the 5′ untranslated region of the PRPF19 mRNA encodes an upstream ORF that is translated in human cells. Inactivation of this upstream ORF reduces the viability of human cells.</p>","PeriodicalId":529,"journal":{"name":"Doklady Biochemistry and Biophysics","volume":"515 1","pages":"41 - 47"},"PeriodicalIF":0.8,"publicationDate":"2024-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11021245/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140108657","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-02-20DOI: 10.1134/S1607672923600422
Qingyan Lu, Qiannan Yan, Xiaojie Li
In the current study the effects of metformin and cyanidin on the immune system and intestinal flora in rats with type-2 diabetes was investigated. The findings showed that metformin or cyanidin treatment considerably reduced the rise in body weight and glucose levels induced by type-2 diabetes. The type-2 diabetic rats’ glucose tolerance was significantly increased by cyanidin administration comparable to that of metformin. Cyanidin administration resulted in a significant reduction in serum cholesterol and low-density lipoprotein (LDL) levels in rats with type-2 diabetes. Treatment with cyanidin significantly increased the ratio of high-density lipoprotein to low-density lipoprotein in type-2 diabetes rats. Cyanidin administration significantly raised the ratio of Firmicutes to Bacteroidetes in the fecal samples of type-2 diabetic rats compared to the model group. In comparison to the model group, it also significantly raised the levels of Lactobacillusintestinalis, Lactobacillus gasseri, and Lactobacillus reuteri in the type-2 diabetes rats. In type-2 diabetes rat fecal samples, the abundance of Christensenellaceae significantly increased while Enterobacteriaceae and Proteobacteria were found to decrease upon cyanidin administration. Furthermore, cyanidin administration to the rats with type-2 diabetes significantly improved the glucose homeostasis. In conclusion, the study demonstrates that cyanidin enhances glucose homeostasis in rats with type-2 diabetes, potentially through controlling intestinal flora. Thus, cyanidin may be looked into more as a possible therapeutic agent for type 2 diabetes.
{"title":"Regulation of Intestinal Flora and Immune Response by Cyanidin Exhibits Protective Effect against Type-2 Diabetes in Rat Model","authors":"Qingyan Lu, Qiannan Yan, Xiaojie Li","doi":"10.1134/S1607672923600422","DOIUrl":"10.1134/S1607672923600422","url":null,"abstract":"<p>In the current study the effects of metformin and cyanidin on the immune system and intestinal flora in rats with type-2 diabetes was investigated. The findings showed that metformin or cyanidin treatment considerably reduced the rise in body weight and glucose levels induced by type-2 diabetes. The type-2 diabetic rats’ glucose tolerance was significantly increased by cyanidin administration comparable to that of metformin. Cyanidin administration resulted in a significant reduction in serum cholesterol and low-density lipoprotein (LDL) levels in rats with type-2 diabetes. Treatment with cyanidin significantly increased the ratio of high-density lipoprotein to low-density lipoprotein in type-2 diabetes rats. Cyanidin administration significantly raised the ratio of <i>Firmicutes</i> to <i>Bacteroidetes</i> in the fecal samples of type-2 diabetic rats compared to the model group. In comparison to the model group, it also significantly raised the levels of <i>Lactobacillus</i> <i>intestinalis</i>, <i>Lactobacillus gasseri</i>, and <i>Lactobacillus reuteri</i> in the type-2 diabetes rats. In type-2 diabetes rat fecal samples, the abundance of <i>Christensenellaceae</i> significantly increased while <i>Enterobacteriaceae</i> and <i>Proteobacteria</i> were found to decrease upon cyanidin administration. Furthermore, cyanidin administration to the rats with type-2 diabetes significantly improved the glucose homeostasis. In conclusion, the study demonstrates that cyanidin enhances glucose homeostasis in rats with type-2 diabetes, potentially through controlling intestinal flora. Thus, cyanidin may be looked into more as a possible therapeutic agent for type 2 diabetes.</p>","PeriodicalId":529,"journal":{"name":"Doklady Biochemistry and Biophysics","volume":"513 1 supplement","pages":"S67 - S74"},"PeriodicalIF":0.8,"publicationDate":"2024-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139911766","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-02-20DOI: 10.1134/S1607672923700709
Y. V. Khramtsov, A. V. Ulasov, T. N. Lupanova, G. P. Georgiev, A. S. Sobolev
Two eukaryotic cell lines, A549 and A431, with stable expression of the nucleocapsid protein (N-protein) of the SARS-CoV-2 virus fused with the red fluorescent protein mRuby3 were obtained. Using microscopy, the volumes of the cytoplasm and nucleus were determined for these cells. Using quantitative immunoblotting techniques, the concentrations of the N-mRuby3 fusion protein in their cytoplasm were assessed. They were 19 and 9 μM for A549 and A431 cells, respectively. Using these concentrations, the initial rate of N-protein degradation in the studied cells was estimated from the decrease in cell fluorescence. In A549 and A431 cells, it was the same (84 nM per hour). The approach of quantitatively describing the degradation process can be applied to analyze the effectiveness of a wide class of antiviral drugs that cause degradation of viral proteins.
{"title":"Quantitative Description of the N-Protein of the SARS-CoV-2 Virus Degradation in Cells Stably Expressing It under the Influence of New Modular Nanotransporters","authors":"Y. V. Khramtsov, A. V. Ulasov, T. N. Lupanova, G. P. Georgiev, A. S. Sobolev","doi":"10.1134/S1607672923700709","DOIUrl":"10.1134/S1607672923700709","url":null,"abstract":"<p>Two eukaryotic cell lines, A549 and A431, with stable expression of the nucleocapsid protein (N-protein) of the SARS-CoV-2 virus fused with the red fluorescent protein mRuby3 were obtained. Using microscopy, the volumes of the cytoplasm and nucleus were determined for these cells. Using quantitative immunoblotting techniques, the concentrations of the N-mRuby3 fusion protein in their cytoplasm were assessed. They were 19 and 9 μM for A549 and A431 cells, respectively. Using these concentrations, the initial rate of N-protein degradation in the studied cells was estimated from the decrease in cell fluorescence. In A549 and A431 cells, it was the same (84 nM per hour). The approach of quantitatively describing the degradation process can be applied to analyze the effectiveness of a wide class of antiviral drugs that cause degradation of viral proteins.</p>","PeriodicalId":529,"journal":{"name":"Doklady Biochemistry and Biophysics","volume":"513 1 supplement","pages":"S63 - S66"},"PeriodicalIF":0.8,"publicationDate":"2024-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139911765","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-02-20DOI: 10.1134/S1607672923700710
Y. V. Khramtsov, A. V. Ulasov, T. N. Lupanova, G. P. Georgiev, A. S. Sobolev
Modular nanotransporters (MNTs) containing an antibody-like molecule, monobody, to the N‑protein of the SARS-CoV-2 virus, as well as an amino acid sequence that recruits the Keap1 E3 ligase (E3BP) were created. This MNT also included a site for cleavage of the E3BP monobody from the MNT in acidic endocytic compartments. It was shown that this cleavage by the endosomal protease cathepsin B leads to a 2.7-fold increase in the affinity of the E3BP monobody for the N-protein. Using A549 cells with transient expression of the N-protein fused with the fluorescent protein mRuby3, it was shown that incubation with MNT leads to a significant decrease in mRuby3 fluorescence. It is assumed that the developed MNTs can serve as a basis for the creation of new antiviral drugs against the SARS-CoV-2 virus.
我们创建了模块化纳米转运体(MNTs),其中含有一种类似于抗体的分子(单体),可与 SARS-CoV-2 病毒的 N 蛋白结合,还含有一种可诱导 Keap1 E3 连接酶(E3BP)的氨基酸序列。这种 MNT 还包括一个用于在酸性内细胞区室中将 E3BP 单体从 MNT 上裂解的位点。研究表明,内泌体蛋白酶 cathepsin B 的这种裂解会使 E3BP 单体对 N 蛋白的亲和力增加 2.7 倍。使用瞬时表达与荧光蛋白 mRuby3 融合的 N 蛋白的 A549 细胞,结果表明与 MNT 一起孵育会导致 mRuby3 荧光显著降低。据推测,所开发的 MNT 可作为开发针对 SARS-CoV-2 病毒的新型抗病毒药物的基础。
{"title":"Modular Nanotransporters Capable of Causing Intracellular Degradation of the N-Protein of the SARS-CoV-2 Virus in A549 Cells with Temporary Expression of This Protein Fused with a Fluorescent Protein mRuby3","authors":"Y. V. Khramtsov, A. V. Ulasov, T. N. Lupanova, G. P. Georgiev, A. S. Sobolev","doi":"10.1134/S1607672923700710","DOIUrl":"10.1134/S1607672923700710","url":null,"abstract":"<p>Modular nanotransporters (MNTs) containing an antibody-like molecule, monobody, to the N‑protein of the SARS-CoV-2 virus, as well as an amino acid sequence that recruits the Keap1 E3 ligase (E3BP) were created. This MNT also included a site for cleavage of the E3BP monobody from the MNT in acidic endocytic compartments. It was shown that this cleavage by the endosomal protease cathepsin B leads to a 2.7-fold increase in the affinity of the E3BP monobody for the N-protein. Using A549 cells with transient expression of the N-protein fused with the fluorescent protein mRuby3, it was shown that incubation with MNT leads to a significant decrease in mRuby3 fluorescence. It is assumed that the developed MNTs can serve as a basis for the creation of new antiviral drugs against the SARS-CoV-2 virus.</p>","PeriodicalId":529,"journal":{"name":"Doklady Biochemistry and Biophysics","volume":"513 1 supplement","pages":"S60 - S62"},"PeriodicalIF":0.8,"publicationDate":"2024-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139911764","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-02-20DOI: 10.1134/S160767292470073X
A. A. Solodovnikov, S. A. Lavrov, A. S. Shatskikh, V. A. Gvozdev
The heterochromatin position effect is manifested in the inactivation of euchromatin genes transferred to heterochromatin. In chromosomal rearrangements, genes located near the new eu-heterochromatin boundary in the rearrangement (cis-inactivation) and, in rare cases, genes of a region of the normal chromosome homologous to the region of the eu-heterochromatin boundary of the chromosome with the rearrangement (trans-inactivation) are subject to inactivation. The In(2)A4 inversion is able to trans-inactivate the UAS-eGFP reporter gene located on the normal chromosome. We knockdown a number of chromatin proteins using temperature-controlled RNA interference and investigated the effect of knockdown on trans-inactivation of the reporter. We found suppression of trans-inactivation by knockdowns of Su(var)2-HP2, a protein that binds to the key heterochromatin protein HP1a, SAYP, a subunit of the chromatin remodelling complex, and Eggless histone methyltransferase (SETDB1), which introduces a H3K9me3 histone mark, recognized by the HP1a protein. The method of studying the effects of gene knockdown on heterochromatin position effects presented in this work is of independent methodological interest.
{"title":"Effects of Chromatin Structure Modifiers on the trans-Acting Heterochromatin Position Effect in Drosophila melanogaster","authors":"A. A. Solodovnikov, S. A. Lavrov, A. S. Shatskikh, V. A. Gvozdev","doi":"10.1134/S160767292470073X","DOIUrl":"10.1134/S160767292470073X","url":null,"abstract":"<p>The heterochromatin position effect is manifested in the inactivation of euchromatin genes transferred to heterochromatin. In chromosomal rearrangements, genes located near the new eu-heterochromatin boundary in the rearrangement (<i>cis</i>-inactivation) and, in rare cases, genes of a region of the normal chromosome homologous to the region of the eu-heterochromatin boundary of the chromosome with the rearrangement (<i>trans</i>-inactivation) are subject to inactivation. The <i>In</i>(<i>2</i>)<i>A4</i> inversion is able to <i>trans</i>-inactivate the <i>UAS-eGFP</i> reporter gene located on the normal chromosome. We knockdown a number of chromatin proteins using temperature-controlled RNA interference and investigated the effect of knockdown on trans-inactivation of the reporter. We found suppression of trans-inactivation by knockdowns of Su(var)2-HP2, a protein that binds to the key heterochromatin protein HP1a, SAYP, a subunit of the chromatin remodelling complex, and Eggless histone methyltransferase (SETDB1), which introduces a H3K9me3 histone mark, recognized by the HP1a protein. The method of studying the effects of gene knockdown on heterochromatin position effects presented in this work is of independent methodological interest.</p>","PeriodicalId":529,"journal":{"name":"Doklady Biochemistry and Biophysics","volume":"513 1 supplement","pages":"S75 - S81"},"PeriodicalIF":0.8,"publicationDate":"2024-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139911762","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}