Phytic acid is the main storage of phosphate in grains of staple crops. As phytic acid is hardly digestible for non-ruminants microbial phytases are used to supplement animal feed to enhance phosphate digestibility. A fungal phytase gene was introduced into barley with the aim of enhancing phosphate digestibility. Transgenic homozygous barley over-expressing fungal phytase phyA showed a 3.3fold increase in mature grain phytase activity. Field trials at two locations in the Czech Republic were conducted in a five-year experiment to test transgene stability and activity under field conditions. Increased phytase activity gradually decreased over the generations showing the most significant drop in the initial years of field trials. Molecular analysis revealed methylation in the coding sequence of the phyA transgene, suggesting transcription gene silencing. On the other hand, herbicide resistance used for selection of transgenic plants was functional over all generations. The feasibility of crossing the transgene into the feeding cultivar Azit was demonstrated with subsequent stabilization of hybrid progeny through androgenesis. Our results indicate that the Azit genetic background tended to reduce phytase activity in mature grains of hybrids. Grain-specific over-expression of fungal phytase driven by an amylase promoter improved phosphate levels during germination. Unfortunately, a malting experiment revealed that phytase over-expression did not significantly improve malting parameters. In fact, the higher nitrogen content in unmalted grain negatively affected the quality of the malt produced from them.

• Klíčová slova
• Transgenic barley, androgenesis, field trials, hybridization, phytase,
• MeSH
• 6-fytasa * genetika   metabolismus   MeSH
• Aspergillus niger * enzymologie   genetika   MeSH
• fosfáty metabolismus   MeSH
• fungální proteiny * genetika   metabolismus   MeSH
• geneticky modifikované rostliny * genetika   metabolismus   MeSH
• ječmen (rod) * genetika   enzymologie   metabolismus   MeSH
• kyselina fytová metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• 6-fytasa * MeSH
• fosfáty MeSH
• fungální proteiny * MeSH
• kyselina fytová MeSH

Concerning the dismal prognosis of chemoresistant patients with epithelial ovarian carcinoma (EOC), we aimed to follow up the findings of a previous whole-exome sequencing study using an orthogonal Sanger sequencing on the same patients and a separate set of 127 EOC patients (N = 177, all fresh frozen tumor samples). We focused on TP53 as a frequently mutated gene relevant for chemosensitivity, included KRAS as an additional therapeutically relevant target, complemented the study with transcript levels of both genes, and compared results with clinical parameters. All variants in TP53 and KRAS detected by exome sequencing were confirmed. KRAS mutated patients had significantly more frequent FIGO stages I or II (p = .002) and other than high-grade serous tumor subtypes (nonHGSCs) (p < .001), which was connected with lower KRAS transcript levels (p = .004). Patients with nonHGSC subtypes had less frequent TP53 mutations (p = .002). Carriers of TP53 variants disrupting the DNA binding loop had significantly longer platinum-free intervals than the rest (p = .037). Tumors bearing nonsense, frameshift, or splice site TP53 variants had a significantly lower TP53 transcript level, while those with missense variants had significantly higher levels than wild types (p < .001). The normalized intratumoral TP53 and KRAS transcript levels were correlated, and patients with co-mutated genes had poorer overall survival than others (p = .015). Protein levels of both genes significantly correlated with their respective transcripts (p = .028 and p = .001, respectively). Our study points to KRAS as a target for future therapy of nonHGSCs and reveals the prognostic value of TP53 variants in the DNA binding loop.

• Klíčová slova
• Epithelial ovarian carcinoma, KRAS, TP53, platinum sensitivity, transcript expression, variant,
• MeSH
• chemorezistence genetika   MeSH
• dospělí MeSH
• epiteliální ovariální karcinom * genetika   farmakoterapie   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mutace MeSH
• nádorový supresorový protein p53 * genetika   metabolismus   MeSH
• nádory vaječníků * genetika   farmakoterapie   patologie   mortalita   MeSH
• platina * terapeutické užití   farmakologie   MeSH
• prognóza MeSH
• protoonkogenní proteiny p21(ras) * genetika   metabolismus   MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• KRAS protein, human MeSH Prohlížeč
• nádorový supresorový protein p53 * MeSH
• platina * MeSH
• protoonkogenní proteiny p21(ras) * MeSH
• TP53 protein, human MeSH Prohlížeč

Insertion sequences (IS) represent mobile genetic elements that have been shown to be associated with bacterial evolution and adaptation due to their effects on genome plasticity. In Bordetella pertussis, the causative agent of whooping cough, the numerous IS elements induce genomic rearrangements and contribute to the diversity of the global B. pertussis population. Previously, we have shown that the majority of IS-specific endogenous promoters induce the synthesis of alternative transcripts and thereby affect the transcriptional landscape of B. pertussis. Here, we describe the regulatory RNA Rfi2, which is transcribed from the Pout promoter of the IS481 gene BP1118 antisense to the adjacent fim2 gene encoding the major serotype 2 fimbrial subunit of B. pertussis. Among the classical bordetellae, Rfi2 is unique to B. pertussis, suggesting its specific role in virulence. We show that Rfi2 RNA attenuates fim2 transcription and, consequently, the production of the Fim2 protein. Interestingly, the mutant that does not produce Rfi2 displayed significantly increased cytotoxicity towards human macrophages compared to the parental strain. This observation suggests that the Rfi2-mediated reduction in cytotoxicity represents an evolutionary adaptation of B. pertussis that fine-tunes its interaction with the human host. Given the immunogenicity of Fim2, we further hypothesize that Rfi2-mediated modulation of Fim2 production contributes to immune evasion. To our knowledge, Rfi2 represents the first functionally characterized IS element-driven antisense RNA that modulates the expression of a virulence gene.

• Klíčová slova
• Bordetella pertussis, antisense RNA, cytotoxicity towards macrophages, fimbriae serotype 2, insertion sequence, modulation of virulence,
• MeSH
• antigeny bakteriální MeSH
• antisense RNA * genetika   metabolismus   MeSH
• bakteriální fimbrie * genetika   metabolismus   MeSH
• Bordetella pertussis * genetika   patogenita   metabolismus   MeSH
• faktory virulence rodu Bordetella genetika   MeSH
• lidé MeSH
• makrofágy mikrobiologie   MeSH
• pertuse mikrobiologie   MeSH
• promotorové oblasti (genetika) MeSH
• proteiny fimbrií * genetika   metabolismus   MeSH
• regulace genové exprese u bakterií * MeSH
• séroskupina MeSH
• transpozibilní elementy DNA * MeSH
• virulence MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigeny bakteriální MeSH
• antisense RNA * MeSH
• faktory virulence rodu Bordetella MeSH
• fim2 protein, Bordetella MeSH Prohlížeč
• proteiny fimbrií * MeSH
• transpozibilní elementy DNA * MeSH

Histones are positively charged proteins found in the chromatin of eukaryotic cells. They regulate gene expression and are required for the organization and packaging of DNA within the nucleus. Histones are extremely conserved, allowing for transcription, replication, and repair. This review delves into their complex structure and function in DNA assembly, their role in nucleosome assembly, and the higher-order chromatin structures they generate. We look at the five different types of histone proteins: H1, H2A, H2B, H3, H4, and their variations. These histones bind with DNA to produce nucleosomes, the basic units of chromatin that are essential for compacting DNA and controlling its accessibility. Their dynamic control of chromatin accessibility has important implications for genomic stability and cellular activities. We elucidate regulatory mechanisms in both normal and pathological situations by investigating their structural features, diverse interaction mechanisms, and chromatin impact. In addition, we discuss the functions of histone post-translational modifications (PTMs) and their significance in various disorders. These alterations, which include methylation, acetylation, phosphorylation, and ubiquitination, are crucial in regulating histone function and chromatin dynamics. We specifically describe and explore the role of changed histones in the evolution of cancer, neurological disorders, sepsis, autoimmune illnesses, and inflammatory conditions. This comprehensive review emphasizes histone's critical role in genomic integrity and their potential as therapeutic targets in various diseases.

• Klíčová slova
• Chromatin, Disease, Gene expression, Genomic stability, Histones, Nucleosomes, Post-translational modifications (PTMs),
• MeSH
• chromatin metabolismus   genetika   MeSH
• DNA * genetika   metabolismus   chemie   MeSH
• histony * metabolismus   genetika   chemie   MeSH
• lidé MeSH
• nádory * metabolismus   genetika   patologie   MeSH
• nukleozomy metabolismus   genetika   MeSH
• posttranslační úpravy proteinů MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• chromatin MeSH
• DNA * MeSH
• histony * MeSH
• nukleozomy MeSH

Disruption of the epidermal barrier contributes to skin disorders such as atopic dermatitis and psoriasis. The aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor, plays a key role in skin homeostasis and immune regulation. While traditionally associated with toxicity, AhR has emerged as a promising therapeutic target, particularly via tryptophan-derived indoles. To support AhR research in a dermatological context, we developed AhaRaCaT, a stable luciferase-based reporter cell line derived from human keratinocytes (HaCaT), enabling the assessment of AhR transcriptional activity in a skin-relevant model. We characterized the inducibility of AhaRaCaT in response to model AhR ligands (TCDD, BaP, FICZ) in dose- and time-dependent assays. Antagonist profiling with MNF, CH223191, GNF, carvone, and jasmone yielded IC50 values over 4- and 24-hour exposures. A panel of indoles previously studied in other models was evaluated for AhR activation, revealing a robust luciferase response at 4 h that declined at 24 h, consistent with trends observed in other cell types. Selected indoles also induced CYP1A1 mRNA expression and reversed cytokine-induced downregulation of filaggrin in HaCaT cells, highlighting their potential in mitigating inflammation-associated skin barrier defects. In summary, the AhaRaCaT cell line offers a sensitive and physiologically relevant tool for studying AhR signaling in skin, with broad applications in toxicology, dermatological research, and the development of AhR-targeted therapies for inflammatory skin diseases.

• Klíčová slova
• Aryl hydrocarbon receptor, FLG, Filaggrin, HaCaT cell line, Indole derivatives, Reporter gene assay,
• MeSH
• buněčné linie keratinocytů HaCaT MeSH
• buněčné linie MeSH
• cytochrom P-450 CYP1A1 genetika   metabolismus   MeSH
• filagriny MeSH
• indoly * farmakologie   toxicita   MeSH
• keratinocyty * účinky léků   metabolismus   MeSH
• kůže * účinky léků   metabolismus   MeSH
• lidé MeSH
• ligandy MeSH
• luciferasy genetika   metabolismus   MeSH
• receptory aromatických uhlovodíků * metabolismus   genetika   agonisté   antagonisté a inhibitory   MeSH
• reportérové geny MeSH
• transkripční faktory bHLH * metabolismus   genetika   agonisté   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• AHR protein, human MeSH Prohlížeč
• CYP1A1 protein, human MeSH Prohlížeč
• cytochrom P-450 CYP1A1 MeSH
• filagriny MeSH
• FLG protein, human MeSH Prohlížeč
• indoly * MeSH
• ligandy MeSH
• luciferasy MeSH
• receptory aromatických uhlovodíků * MeSH
• transkripční faktory bHLH * MeSH

The retinoic acid-inducible gene-I (RIG-I) signaling is crucial for cell-intrinsic innate antiviral immunity. Upon cytosolic detection of virus-associated RNA, it triggers a cascade inducing production of potent cytokines, mainly type I and III interferons (IFNs). While effective, dysregulated responses can harm the host, requiring tight pathway control. Here, we performed a comprehensive, systematic siRNA-based high-throughput screen across 616 established and putative E3 ubiquitin ligases for their impact on RIG-I signaling. We employed a fluorescence-based live-cell imaging assay in A549 cells to monitor nuclear translocation of IRF3 and NF-κB, two key transcription factors downstream of RIG-I. Candidate genes were validated in an orthogonal secondary screen, assessing their impact on the functional antiviral response to a Rift Valley Fever reporter virus. Fourteen hits showed consistent effects on RIG-I signaling across both screens. These genes were further validated and characterized by assessing IFN-β promoter reporter activity and IFNB1 mRNA levels upon dsRNA transfection. TRIM48 emerged as a highly robust negative regulator. Overexpression of TRIM48 suppressed RIG-I-mediated activation of IRF3 and NF-κB, reduced IFN and IFN-stimulated gene expression, and enhanced viral replication. Conversely, TRIM48 deficiency enhanced RIG-I signaling and inhibited viral replication. Notably, TRIM48 acts as an induced feedback regulator upon infection, and its effect depended on its enzymatic ubiquitin ligase activity. Our high-throughput screen provides an unbiased assessment of close to all E3 ubiquitin ligases for their regulatory effect in RIG-I signaling, and identified several interesting candidates for further investigation. TRIM48 was established as a negative feedback regulator of the RIG-I pathway.

• Klíčová slova
• E3 ubiquitin ligases, Innate antiviral immunity, RIG-I signaling, TRIM48, siRNA screening,
• MeSH
• buňky A549 MeSH
• DEAD box protein 58 * metabolismus   MeSH
• HEK293 buňky MeSH
• interferon beta genetika   metabolismus   MeSH
• interferonový regulační faktor 3 metabolismus   MeSH
• lidé MeSH
• NF-kappa B metabolismus   MeSH
• receptory imunologické MeSH
• rychlé screeningové testy MeSH
• signální transdukce * MeSH
• TRIM protein MeSH
• ubikvitinligasy * metabolismus   genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• DEAD box protein 58 * MeSH
• interferon beta MeSH
• interferonový regulační faktor 3 MeSH
• IRF3 protein, human MeSH Prohlížeč
• NF-kappa B MeSH
• receptory imunologické MeSH
• RIGI protein, human MeSH Prohlížeč
• TRIM protein MeSH
• ubikvitinligasy * MeSH

While cytokinin (CK) can delay natural leaf senescence, its effects on abiotic stress accelerated leaf senescence are less studied. Here we show N-conjugated trans-zeatin CK forms (tZ7G and tZ9G, or tZNGs) have the ability to delay salt stress senescence. Using a modified dark-induced senescence bioassay with Arabidopsis leaves, exogenous salt treatment accelerated leaf senescence as measured by lower photosystem II efficiency (Fv/Fm) and chlorophyll content. tZNGs were able to delay these parameters at concentrations as low as 10 nM similar to tZ, indicating that tZ7G and tZ9G can function in delaying salt accelerated senescence (SAS). To better understand physiological effects regulating tZNG delay of senescence, transcriptomics, proteomics, as well as CK measurements were examined. Salt treatment has strong transcriptome and proteome effects in accelerating senescence and reducing overall CK levels. Exogenous CK treatments could be quickly detected from changes seen in endogenous CK measurements, where each CK has a distinct profile contributing to transcript/protein alterations. Interestingly, transcriptomics show tZNGs are primarily responsive at later stages of salt senescence, in contrast to an immediate and continual response of tZ treatment. Known CK-regulated genes are induced by tZNGs and tZ, as corroborated by ARR:GUS reporter lines. Differences between tZNGs and tZ DEGs were revealed by WGCNA that included salt and CK specifically gene modules. In contrast, proteomic analysis revealed unique, but similar numbers of tZNG compared to tZ DAPs across senescence. GO term analysis of tZNG DEGs and DAPs showed enrichment of senescence, chloroplast, and CK signaling. Together this indicates tZNGs function as active CK forms in delaying salt accelerated leaf senescence.

• Klíčová slova
• Cytokinin level, Cytokinin-N-Glucosides, Leaf senescence, Proteome, Salt-stress, Transcriptome, WGCNA, trans-Zeatin,
• MeSH
• Arabidopsis * účinky léků   metabolismus   fyziologie   genetika   MeSH
• chlorid sodný * farmakologie   MeSH
• chlorofyl metabolismus   MeSH
• cytokininy metabolismus   MeSH
• glukosidy * farmakologie   metabolismus   MeSH
• listy rostlin * účinky léků   metabolismus   fyziologie   MeSH
• regulace genové exprese u rostlin účinky léků   MeSH
• senescence rostlin * účinky léků   MeSH
• zeatin * farmakologie   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• chlorid sodný * MeSH
• chlorofyl MeSH
• cytokininy MeSH
• glukosidy * MeSH
• zeatin * MeSH

Linking meta-omics and biogeochemistry approaches in soils has remained challenging. This study evaluates the use of an internal RNA extraction standard and its potential for making quantitative estimates of a given microbial community size (biomass) in soil metatranscriptomics. We evaluate commonly used laboratory protocols for RNA processing, metatranscriptomic sequencing and quantitative reverse transcription polymerase chain reaction (qRT-PCR). Metatranscriptomic profiles from soil samples were generated using two library preparation protocols and prepared in triplicates. RNA extracted from pure cultures of Saccharolobus solfataricus was added to the samples as an internal nucleic acid extraction standard (NAEstd). RNA reads originating from NAEstd were identified with a 99.9% accuracy. A remarkable replication consistency between triplicates was seen (average Bray-Curtis dissimilarity 0.03 ± 0.02), in addition to a clear library preparation bias. Nevertheless, the between-sample pattern was not affected by library type. Estimates of 16S rRNA transcript abundance derived from qRT-PCR experiments, NAEstd and a previously published quantification method of metatranscriptomics (hereafter qMeTra) were compared with microbial biomass carbon (MBC) and nitrogen (MBN) extracts. The derived biomass estimates differed by orders of magnitude. While most estimates were significantly correlated with each other, no correlation was observed between NAEstd and MBC extracts. We discuss how simultaneous changes in community size and the soils nucleic acid retention strength might hamper accurate biomass estimation. Adding NAEstd has the potential to shed important light on nucleic acid retention in the substance matrix (e.g., soil) during extraction.

• Klíčová slova
• RNA, biomass estimates, extraction standard, metatranscriptomics, quantitative transcriptomics,
• MeSH
• Bacteria * genetika   klasifikace   MeSH
• metagenomika * metody   MeSH
• mikrobiota * MeSH
• půda chemie   MeSH
• půdní mikrobiologie * MeSH
• RNA ribozomální 16S genetika   MeSH
• stanovení celkové genové exprese * metody   MeSH
• transkriptom MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• půda MeSH
• RNA ribozomální 16S MeSH

This perspective highlights the emerging significance of noncanonical nucleic acid structures-such as G-quadruplexes, Z-DNA/Z-RNA, and cruciforms-in viral genomes. Once considered structural oddities, these motifs are now recognized as critical regulators of viral replication, transcription, genome stability, and host-pathogen interactions. Despite mounting evidence of their functional relevance and therapeutic potential, these structures remain largely overlooked in virology and antiviral drug development. Their unique conformations offer highly specific molecular targets, with several small molecules already demonstrating the ability to modulate viral gene expression by stabilizing or destabilizing these motifs. The persistent underestimation of non-B DNA/RNA structures represents a missed opportunity in the fight against viral diseases. By synthesizing recent discoveries and emphasizing their biological and pharmacological promise, we aim to elevate awareness and catalyze interdisciplinary research. Harnessing the structural diversity of viral genomes could unlock novel antiviral strategies with high specificity and minimal off-target effects.

• Klíčová slova
• DNA structure, G-quadruplex, Z-DNA, cruciform, targeting viruses,
• Publikační typ
• časopisecké články MeSH

The nuclear envelope is subtended in most eukaryotes by a proteinaceous lamina, a network that has been recognised since the 1950s. Originally considered as a simple structural support, it is now clear that the lamina can be highly dynamic and participates in a multitude of functions, including transcriptional and epigenetic regulation, definition of chromatin domains, genome stability and the positioning of nuclear pore complexes. The major protein components of the lamina in metazoans are ~60 kDa lamins, which assemble to form fibres and a network and are regulated by multiple mechanisms. Despite a broad taxonomic distribution beyond Metazoa, lamins are not universal and, in at least three major lineages, are absent, specifically fungi, plants and kinetoplastid protists. The latter two possess lineage-specific lamin analogues, the CRWN and NUP-1/NUP-2 proteins, respectively. Here we discuss and compare the kinetoplastid and plant lamina, their origins, components and functions and spectacular examples of convergent evolution.

• Klíčová slova
• CRWN proteins, Eukaryogenesis, Evolution, Gene expression, Lamina, Nuclear pore complex, Nucleus, Plants, Trypanosomes,
• MeSH
• jaderná lamina * metabolismus   MeSH
• laminy * metabolismus   genetika   MeSH
• lidé MeSH
• rostliny metabolismus   genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• laminy * MeSH