Doklady Biochemistry and Biophysics最新文献

{"title":"Investigation of the Functional Role of the Conserved Sequence at the 5'-End of the Fourth Intron of the mod(mdg4) Gene in Trans-Splicing in Drosophila melanogaster","authors":"Iu. V. Soldatova,&nbsp;O. Beginyazova,&nbsp; P. G. Georgiev,&nbsp;M. V. Tikhonov","doi":"10.1134/S1607672924601410","DOIUrl":"10.1134/S1607672924601410","url":null,"abstract":"<p>Alternative splicing is an important mechanism that provides genetic diversity of proteins. Unique loci have been identified in <i>Drosophila melanogaster</i>, where mRNA diversity arises as a result of <i>trans</i>-splicing—a process in which exons from different pre-mRNAs are joined together. The <i>trans</i>-splicing in the <i>mod(mdg4)</i> locus, which encodes more than 31 isoforms, has been studied in detail. Important elements for this process include previously described conserved sequences in the fourth intron. The aim of this study is to further characterize the conserved motifs of the fourth intron, specifically the element at the 5'-end of the intron. Using model transgenic lines, it has been shown that introduced changes in the sequence of the studied element lead to a disruption of <i>trans</i>-splicing. In contrast, similar changes in the endogenous locus did not result in a disruption of <i>trans</i>-splicing. Thus, the conserved element plays a role in <i>trans</i>-splicing but is not critical.</p>","PeriodicalId":529,"journal":{"name":"Doklady Biochemistry and Biophysics","volume":"521 1","pages":"169 - 173"},"PeriodicalIF":0.8,"publicationDate":"2025-04-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143949625","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}

{"title":"Identification of a Highly Conserved Region Critical for the Functionality of the Cp190 Protein in Drosophila melanogaster","authors":"L. S. Melnikova,&nbsp;K. I. Balagurov,&nbsp; P. G. Georgiev,&nbsp;A. K. Golovnin","doi":"10.1134/S1607672924601434","DOIUrl":"10.1134/S1607672924601434","url":null,"abstract":"<p>The CP190 protein binds to both housekeeping gene promoters and insulator/boundary elements and plays a critical role in their activity. The aim of this work was to study the effect of deletions in highly conserved regions of the CP190 protein on its functionality. It was shown that deletion of the sequence from 664 to 700 aa leads to a lethal phenotype. Thus, a new region was identified in CP190 that plays an important role in the interaction of the CP190 protein with as yet unidentified partner proteins, stabilization of protein complexes formed by CP190, and their recruitment to chromatin.</p>","PeriodicalId":529,"journal":{"name":"Doklady Biochemistry and Biophysics","volume":"521 1","pages":"178 - 182"},"PeriodicalIF":0.8,"publicationDate":"2025-04-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143949624","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}

{"title":"The bxd PRE from Bithorax Complex of Drosophila melanogaster Has Weak Insulator Activity","authors":"A. N. Ibragimov,&nbsp;O. V. Kyrchanova,&nbsp;Y. E. Vorontsova,&nbsp;E. N. Kozlov,&nbsp;V. O. Dubrovskaya,&nbsp; P. G. Georgiev","doi":"10.1134/S1607672924601513","DOIUrl":"10.1134/S1607672924601513","url":null,"abstract":"<p>The autonomous functioning of regulatory domains in the Bithorax complex (BX-C) of <i>Drosophila melanogaster</i> is maintained by boundaries (insulators) that prevent inappropriate enhancer–promoter interactions. Polycomb response elements (PREs) maintain epigenetic memory within regulatory domains and are often located near insulators, enhancing their blocking activity. The <i>bxd</i> PRE is a well-characterized silencer that regulates the <i>bxd</i>/<i>pbx</i> domain of the <i>Ubx</i> gene in the first abdominal segment. We used a boundary replacement strategy to assess the insulator activity of the <i>bxd</i> PRE. Substituting the <i>bxd</i> PRE for the <i>Fab-</i>7 boundary, which separates the <i>Abd-B</i> regulatory domains in the BX-C, revealed that the <i>bxd</i> PRE has weak insulator activity.</p>","PeriodicalId":529,"journal":{"name":"Doklady Biochemistry and Biophysics","volume":"521 1","pages":"183 - 186"},"PeriodicalIF":0.8,"publicationDate":"2025-04-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143949627","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}

{"title":"Functional Role of Bap170 Domains in Enhancer-Dependent Gene Activity in Drosophila melanogaster","authors":"V. K. Chmykhalo,&nbsp;D. Amendola,&nbsp;Y. V. Shidlovskii,&nbsp;L. A. Lebedeva,&nbsp; P. Schedl,&nbsp;E. Giordano","doi":"10.1134/S1607672924601409","DOIUrl":"10.1134/S1607672924601409","url":null,"abstract":"<p>The Bap170 subunit of the SWI/SNF chromatin remodeler exhibits activator functions when artificially recruited to the <i>LacZ</i> reporter promoter in enhancer-dependent transcription. In this study, the functional significance of Bap170 protein domains in reporter activation was analyzed. Deletion of the ARID domain does not reduce Bap170 activity. Increased expression of the <i>LacZ</i> reporter was observed in the form of Bap170 without a region that includes LXXLL motifs. Deletions of the central (RFX domain and IDRs) and C-terminal region (zinc fingers) of Bap170 lead to a significant decrease in transgene expression. Apparently, these regions of Bap170 are critical for the function of this protein in enhancer-dependent transcription.</p>","PeriodicalId":529,"journal":{"name":"Doklady Biochemistry and Biophysics","volume":"520 1","pages":"152 - 155"},"PeriodicalIF":0.8,"publicationDate":"2025-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143078299","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}

{"title":"Evolutionarily Conserved DHX9/MLE Helicase Is Involved in the Regulation of Its Own mRNA Expression Level in Drosophila melanogaster","authors":"I. A. Zolin,&nbsp; S. G. Georgieva,&nbsp;J. V. Nikolenko","doi":"10.1134/S1607672924601239","DOIUrl":"10.1134/S1607672924601239","url":null,"abstract":"<p>The MLE helicase of <i>D. melanogaster</i>, like its ortholog DHX9 in mammals, is involved in a wide range of processes related to the regulation of gene expression. In the present study, we investigated the impact of the <i>mle[9]</i> mutation on its own mRNA expression level. It was shown that in addition to the previously described deletion in the catalytic domain of the protein, which impairs its helicase activity, the <i>mle[9]</i> mutation contains an additional small deletion in the C-terminal domain. In the <i>mle[9]</i> mutation background, there was a threefold increase in the expression of the main transcript of the <i>mle</i> gene encoding the full-length protein. Binding of MLE to chromatin at the coding region and promoters of the <i>mle</i> gene and nearby enhancers was analyzed. To exclude the influence of dosage compensation, experiments were performed on females. The data obtained indicate the role of MLE in specific regulation of its own mRNA expression level in vivo at the adult stage.</p>","PeriodicalId":529,"journal":{"name":"Doklady Biochemistry and Biophysics","volume":"520 1","pages":"1 - 5"},"PeriodicalIF":0.8,"publicationDate":"2025-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143078297","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}

{"title":"An Investigation of the Conservatism of Sequences Defining trans-Splicing in the mod(mdg4) Locus across Drosophila and Silkworm Species","authors":"O. Beginyazova,&nbsp;I. V. Soldatova,&nbsp; P. G. Georgiev,&nbsp;M. V. Tikhonov","doi":"10.1134/S1607672924601021","DOIUrl":"10.1134/S1607672924601021","url":null,"abstract":"<p>Splicing, a key step in mRNA maturation, plays a crucial role in the regulation of eukaryotic gene expression. The formation of chimeric mRNAs from two transcripts during splicing is typically a prohibited process; however, in insects, <i>trans</i>-splicing is a primary mechanism for increasing protein diversity for several loci. The aim of this study is to investigate the evolutionary conservativeness of sequences responsible for <i>trans</i>-splicing in the <i>mod(mdg4)</i> locus among species from the Drosophilidae family (order Diptera) and the silkworm (<i>Bombyx mori</i>), which belongs to the order Lepidoptera. Using model transgenic lines, it was shown that sequences from distant <i>Drosophila</i> species retain the ability to support <i>trans</i>-splicing in <i>D. melanogaster</i>. In contrast, analogous sequences in <i>Bombyx mori</i> do not support <i>trans</i>-splicing. Thus, the RNA motifs and their binding hypothetical protein factors, defining <i>trans</i>-splicing, remain conserved among the Drosophilidae group, but have functionally diverged between Diptera and Lepidoptera.</p>","PeriodicalId":529,"journal":{"name":"Doklady Biochemistry and Biophysics","volume":"520 1","pages":"144 - 147"},"PeriodicalIF":0.8,"publicationDate":"2025-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143078295","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}

{"title":"GALA3-Containing Modular Nanotransporters Are Capable of Delivering Keap1 Monobody to Target Cells and Inhibiting the Formation of Reactive Oxygen Species in the Cells","authors":"Y. V. Khramtsov,&nbsp;E. S. Bunin,&nbsp;A. V. Ulasov,&nbsp;T. N. Lupanova,&nbsp;G. P. Georgiev,&nbsp;A. S. Sobolev","doi":"10.1134/S1607672924601252","DOIUrl":"10.1134/S1607672924601252","url":null,"abstract":"<p>In the previously created modular nanotransporter (MNT) capable of delivering a monobody to Keap1 into the cytosol, the endosomolytic module, translocation domain of diphtheria toxin (DTox), was replaced by the endosomolytic peptide GALA3. It was found that this substitution more than doubles the lifetime of MNT in the blood. Using confocal microscopy, it was shown that MNT with GALA3 was internalized into AML12 cells mainly due to binding to the epidermal growth factor receptor, and is also able to exit from endosomes into the cytosol. Using cellular thermal shift assay, it was shown that MNT with GALA3 and MNT with DTox are equally effective in disrupting the formation of the Nrf2 complex with Keap1, which led to similar protection of AML12 cells from the action of hydrogen peroxide. The obtained results allow not only optimizing the systemic use of MNT, but can also serve as a basis for creating agents aimed at treating diseases associated with oxidative stress.</p>","PeriodicalId":529,"journal":{"name":"Doklady Biochemistry and Biophysics","volume":"520 1","pages":"148 - 151"},"PeriodicalIF":0.8,"publicationDate":"2025-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143078303","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}

{"title":"New N6-Substituted Adenine Derivatives with High Antiviral Activity against RNA-Containing Viruses","authors":"A. A. Zenchenko,&nbsp;Yu. D. Semenova,&nbsp;E. R. Naberezhnaya,&nbsp;Ya. D. Gumennaya,&nbsp;A. V. Lipatova,&nbsp;V. E. Oslovsky","doi":"10.1134/S1607672924600787","DOIUrl":"10.1134/S1607672924600787","url":null,"abstract":"<p>In this work, two new compounds, <i>N</i>⁶-(4,5-dimethoxyphenyl)adenine and <i>N</i>⁶-(3,5-di-trifluoromethylphenyl)adenine, with a broad range of antiviral activity against RNA viruses were identified. We showed that these compounds exhibit pronounced antiviral activity against human poliovirus types 1, 2, and 3, belonging to enterovirus C species. Both compounds also demonstrated pronounced antiviral activity against Coxsackie viruses B3, B5, and B6, belonging to enterovirus B species. In addition, the compounds demonstrated antiviral activity against Newcastle disease virus, which belongs to the paramyxovirus genus. The compounds discovered in this work can subsequently serve as prototypes for the development of new antiviral drugs against epidemiologically significant human RNA viruses.</p>","PeriodicalId":529,"journal":{"name":"Doklady Biochemistry and Biophysics","volume":"520 1","pages":"38 - 41"},"PeriodicalIF":0.8,"publicationDate":"2025-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143021358","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}

{"title":"Low-molecular-weight Ligand of the Thyroid-stimulating Hormone Receptor with the Activity of a Partial Agonist and a Negative Allosteric Modulator","authors":"K. V. Derkach,&nbsp;E. A. Didenko,&nbsp;V. N. Sorokoumov,&nbsp;I. O. Zakharova,&nbsp;A. O. Shpakov","doi":"10.1134/S1607672924600799","DOIUrl":"10.1134/S1607672924600799","url":null,"abstract":"<p>Graves’ disease is caused by overactivation of the thyroid-stimulating hormone receptor (TSHR). One approach for its treatment may be the use of negative allosteric modulators (NAM) of TSHR, which normalize TSHR activity and do not cause thyroid hormone (TH) deficiency. The aim of the work was to study the effect of a new compound 5-amino-4-(4-bromophenyl)-2-(methylthio)thieno[2,3-d]pyrimidine-6-carboxylic acid <i>N</i>-<i>tert</i>-butylamide (TPY4) on the basal and TSH-stimulated TH production in cultured FRTL-5 thyrocytes and on basal and thyrotropin-releasing hormone (TRH)-stimulated TH levels in the blood of rats. TPY4 stimulated TH production by thyrocytes and increased TH levels when administered intraperitoneally and orally in rats. It also decreased the TSH-stimulated TH production in thyrocytes and the TRH-stimulated TH levels in rats. Thus, TPY4 is the first known allosteric regulator of TSHR, combining the properties of NAM and a partial agonist, and can be considered as a prototype of drugs for the treatment of Graves’ disease.</p>","PeriodicalId":529,"journal":{"name":"Doklady Biochemistry and Biophysics","volume":"520 1","pages":"53 - 57"},"PeriodicalIF":0.8,"publicationDate":"2025-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143021392","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}

{"title":"Method for Testing Drugs Belonging to Substrates and Inhibitors of the Transporter Protein BCRP on CACO-2 Cells","authors":"Yu. S. Tranova,&nbsp;A. A. Slepnev,&nbsp;I. V. Chernykh,&nbsp;A. V. Shchulkin,&nbsp;P. Yu. Mylnikov,&nbsp;N. M. Popova,&nbsp;M. I. Povetko,&nbsp;E. N. Yakusheva","doi":"10.1134/S1607672924701266","DOIUrl":"10.1134/S1607672924701266","url":null,"abstract":"<p>Introduction. Breast cancer resistance protein (BCRP) is an efflux membrane transporter that controls the pharmacokinetics of a large number of drugs. Its activity may change when taking some endo- and exogenous substances, thus making it a link in drug interactions. \u0000<b>Aim.</b> The aim of the study was to develop a methodology for testing drugs for belonging to BCRP substrates and inhibitors in vitro. \u0000<b>Materials and methods.</b> The work was performed on Caco-2 cells overexpressing BCRP, the cultivation was performed in a transwell system consisting of the apical and basolateral chambers. Cells were seeded at the bottom of the apical chamber, which is a semi-permeable membrane. Primarily, the transport of BCRP substrates—methotrexate, mitoxantrone, and quercetin—was evaluated in the concentration range of 1, 5, 10, and 50 μM in the direction from the basal chamber to the apical one (Papp b-a) and in the opposite direction (Papp a-b). The Papp b-a/Papp a-b ratio more than 2 characterizes the involvement of transporter proteins in the transcellular transport of substances. To confirm the involvement of BCRP in their transport, an experiment was carried out with the addition of a transporter inhibitor, reserpine, at a concentration of 50 μM to the transport medium. The concentration of substrates in the chambers was analyzed by HPLC–MS/MS. \u0000<b>Results.</b> After the addition of methotrexate (1 μM), mitoxantrone (1 μM), and quercetin (1–10 μM) to the apical or basolateral chambers of the transwell system, their content in the recipient chamber was not detected. \u0000At methotrexate concentration of 5 μM, the Papp b-a/Papp a-b ratio was 3.38 ± 0.08, which indicates the involvement of transporters in its transfer. When methotrexate was added to the donor chamber at concentrations of 10 and 50 μM, the Papp b-a/Papp a-b ratio decreased to values below 2. \u0000At mitoxantrone concentration of 5 μM, the Papp b-a/Papp a-b ratio was 2.72 ± 0.16. An increase in the concentration to 10 μM led to an increase in the Papp b-a/Papp a-b ratio to 6.18 ± 0.08. At the substance concentration of 50 μM, the index decreased but remained above 2. \u0000At the quercetin concentration of 50 µM, the Papp b-a/Papp ratio was below 2. \u0000Reserpine reduced the Papp b-a/Papp a-b ratio of methotrexate 3.31 times (<i>p</i> = 0.0002), which indicates the elimination of asymmetry in the transport of the substance. At a mitoxantrone concentration of 10 µM, reserpine reduced its Papp b-a/Papp a-b ratio 3.36 times (<i>p</i> &lt; 0.0001). These results indicate the involvement of BCRP in the control of the transfer of both substances through the cellular monolayer. \u0000<b>Conclusions.</b> A method of testing drugs for affiliation to BCRP substrates and inhibitors using methotrexate (5 μM) and mitoxantrone (10 μM) as marker substrates and reserpine (50 μM) as inhibitor was developed and tested on Caco-2 cells.</p>","PeriodicalId":529,"journal":{"name":"Doklady Biochemistry and Biophysics","volume":"520 1","pages":"89 - 95"},"PeriodicalIF":0.8,"publicationDate":"2025-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143021354","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}