Tardigrades are microscopic ecdysozoans that can withstand extreme environmental conditions. Several tardigrade species undergo reversible morphological transformations and enter into cryptobiosis, which helps them to survive periods of unfavorable environmental conditions. However, the underlying molecular mechanisms of cryptobiosis are mostly unknown. Tubulins are evolutionarily conserved components of the microtubule cytoskeleton that are crucial in many cellular processes. We hypothesize that microtubules are necessary for the morphological changes associated with successful cryptobiosis. The molecular composition of the microtubule cytoskeleton in tardigrades is unknown. Therefore, we analyzed and characterized tardigrade tubulins and identified 79 tardigrade tubulin sequences in eight taxa. We found three α-, seven β-, one γ-, and one ε-tubulin isoform. To verify in silico identified tardigrade tubulins, we also isolated and sequenced nine out of ten predicted Hypsibius exemplaris tubulins. All tardigrade tubulins were localized as expected when overexpressed in mammalian cultured cells: to the microtubules or to the centrosomes. The presence of a functional ε-tubulin, clearly localized to centrioles, is attractive from a phylogenetic point of view. Although the phylogenetically close Nematoda lost their δ- and ε-tubulins, some groups of Arthropoda still possess them. Thus, our data support the current placement of tardigrades into the Panarthropoda clade.
- MeSH
- fylogeneze * MeSH
- Tardigrada * klasifikace MeSH
- tubulin genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Arthroderma is the most diverse genus among dermatophytes encompassing species occurring in soil, caves, animal burrows, clinical material and other environments. In this study, we collected ex-type, reference and authentic strains of all currently accepted Arthroderma species and generated sequences of three highly variable loci (ITS rDNA, β-tubulin, and translation elongation factor 1-α). The number of accepted species was expanded to 27. One novel species, A. melbournense (ex-type strain CCF 6162T = CBS 145858T), is described. This species was isolated from toenail dust collected by a podiatrist in Melbourne, during an epidemiological study of four geographical regions of Eastern Australia. Trichophyton terrestre, Chrysosporium magnisporum, and Chrysosporium oceanitis are transferred to Arthroderma. Typification is provided for T. terrestre that is not conspecific with any of the supposed biological species from the former T. terrestre complex, that is, A. insingulare, A. lenticulare and A. quadrifidum. A multi-gene phylogeny and reference sequences provided in this study should serve as a basis for future phylogenetic studies and facilitate species identification in practice.LAY ABSTRACT: The genus Arthroderma encompasses geophilic dermatophyte species that infrequently cause human and animal superficial infections. Reference sequences from three genetic loci were generated for all currently accepted Arthroderma species and phylogeny was constructed. Several taxonomic novelties are introduced. The newly provided data will facilitate species identification and future taxonomic studies.
- MeSH
- Arthrodermataceae klasifikace genetika MeSH
- DNA fungální genetika MeSH
- elongační faktor 1 genetika MeSH
- fylogeneze * MeSH
- geny hub genetika MeSH
- lidé MeSH
- mezerníky ribozomální DNA genetika MeSH
- Microsporum klasifikace genetika MeSH
- Trichophyton klasifikace genetika MeSH
- tubulin genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Austrálie MeSH
Fluorescent nanodiamonds (NDs) coated with therapeutics and cell-targeting structures serve as effective tools for drug delivery. However, NDs circulating in blood can eventually interact with the blood-brain barrier, resulting in undesired pathology. Here, we aimed to detect interaction between NDs and adult brain tissue. First, we cultured neuronal tissue with ND ex vivo and studied cell prosperity, regeneration, cytokine secretion, and nanodiamond uptake. Then, we applied NDs systemically into C57BL/6 animals and assessed accumulation of nanodiamonds in brain tissue and cytokine response. We found that only non-neuronal cells internalized coated nanodiamonds and responded by excretion of interleukin-6 and interferon-γ. Cells of neuronal origin expressing tubulin beta-III did not internalize any NDs. Once we applied coated NDs intravenously, we found no presence of NDs in the adult cortex but observed transient release of interleukin-1α. We conclude that specialized adult neuronal cells do not internalize plain or coated NDs. However, coated nanodiamonds interact with non-neuronal cells present within the cortex tissue. Moreover, the coated NDs do not cross the blood-brain barrier but they interact with adjacent barrier cells and trigger a temporary cytokine response. This study represents the first report concerning interaction of NDs with adult brain tissue.
- MeSH
- biokompatibilní potahované materiály chemie farmakologie MeSH
- interferon gama metabolismus MeSH
- interleukin-6 metabolismus MeSH
- kultivované buňky MeSH
- mozek patologie MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- nanodiamanty chemie toxicita MeSH
- neurony cytologie účinky léků metabolismus MeSH
- regenerace účinky léků MeSH
- tubulin genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Highly conserved α- and β-tubulin heterodimers assemble into dynamic microtubules and perform multiple important cellular functions such as structural support, pathway for transport and force generation in cell division. Tubulin exists in different forms of isotypes expressed by specific genes with spatially- and temporally-regulated expression levels. Some tubulin isotypes are differentially expressed in normal and neoplastic cells, providing a basis for cancer chemotherapy drug development. Moreover, specific tubulin isotypes are overexpressed and localized in the nuclei of cancer cells and/or show bioenergetic functions through the regulation of the permeability of mitochondrial ion channels. It has also become clear that tubulin isotypes are involved in multiple cellular functions without being incorporated into microtubule structures. Understanding the mutations of tubulin isotypes specifically expressed in tumors and their post-translational modifications might help to identify precise molecular targets for the design of novel anti-microtubular drugs. Knowledge of tubulin mutations present in tubulinopathies brings into focus cellular functions of tubulin in brain pathologies such as Alzheimer's disease. Uncovering signaling pathways which affect tubulin functions during antigen-mediated activation of mast cells presents a major challenge in developing new strategies for the treatment of inflammatory and allergic diseases. γ-tubulin, a conserved member of the eukaryotic tubulin superfamily specialized for microtubule nucleation is a target of cell cycle and stress signaling. Besides its microtubule nucleation role, γ-tubulin functions in nuclear and cell cycle related processes. This special issue "Tubulin: Structure, Functions and Roles in Disease" contains eight articles, five of which are original research papers and three are review papers that cover diverse areas of tubulin biology and functions under normal and pathological conditions.
- MeSH
- Alzheimerova nemoc genetika metabolismus patologie MeSH
- lidé MeSH
- mikrotubuly genetika metabolismus patologie MeSH
- mutace MeSH
- nádorové proteiny genetika metabolismus MeSH
- nádory genetika metabolismus MeSH
- protein - isoformy MeSH
- tubulin genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
- úvodní články MeSH
- úvodníky MeSH
Fungi under the order Ophiostomatales (Ascomycota) are known to associate with various species of bark beetles (Coleoptera: Curculionidae: Scolytinae). In addition this group of fungi contains many taxa that can impart blue-stain on sapwood and some are important tree pathogens. A recent survey that focussed on the diversity of the Ophiostomatales in the forest ecosystems of the Czech Republic and Poland uncovered four putative new species. Phylogenetic analyses of four gene regions (ITS1-5.8S-ITS2 region, ß-tubulin, calmodulin, and translation elongation factor 1-α) indicated that these four species are members of the genus Ophiostoma. All four newly described species can be distinguished from each other and from closely related species based on DNA sequence comparisons, morphological characters, growth rates, and their insect associations. Based on this study four new taxa can be circumscribed and the following names are provided: Ophiostoma pityokteinis sp. nov., Ophiostoma rufum sp. nov., Ophiostoma solheimii sp. nov., and Ophiostoma taphrorychi sp. nov. O. rufum sp. nov. is a member of the Ophiostoma piceae species complex, while O. pityokteinis sp. nov. resides in a discrete lineage within Ophiostoma s. stricto. O. taphrorychi sp. nov. together with O. distortum formed a well-supported clade in Ophiostoma s. stricto close to O. pityokteinis sp. nov. O. solheimii sp. nov. groups within a currently undefined lineage A, which also includes Ophiostoma grandicarpum and Ophiostoma microsporum. This study highlights the need for more intensive surveys that should include additional countries of Central Europe, insect vectors and host tree species in order to elucidate Ophiostoma species diversity in this region.
- MeSH
- cévnaté rostliny parazitologie MeSH
- dřevo parazitologie MeSH
- elongační faktor 1 genetika MeSH
- fylogeneze * MeSH
- kalmodulin genetika MeSH
- mezerníky ribozomální DNA genetika MeSH
- mykologické určovací techniky MeSH
- nosatcovití růst a vývoj mikrobiologie MeSH
- Ophiostoma klasifikace genetika izolace a purifikace fyziologie MeSH
- RNA ribozomální 5.8S genetika MeSH
- sekvenční analýza DNA MeSH
- tubulin genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
- Polsko MeSH
De novo heterozygous missense variants in the γ-tubulin gene TUBG1 have been linked to human malformations of cortical development associated with intellectual disability and epilepsy. Here, we investigated through in-utero electroporation and in-vivo studies, how four of these variants affect cortical development. We show that TUBG1 mutants affect neuronal positioning, disrupting the locomotion of new-born neurons but without affecting progenitors' proliferation. We further demonstrate that pathogenic TUBG1 variants are linked to reduced microtubule dynamics but without major structural nor functional centrosome defects in subject-derived fibroblasts. Additionally, we developed a knock-in Tubg1Y92C/+ mouse model and assessed consequences of the mutation. Although centrosomal positioning in bipolar neurons is correct, they fail to initiate locomotion. Furthermore, Tubg1Y92C/+ animals show neuroanatomical and behavioral defects and increased epileptic cortical activity. We show that Tubg1Y92C/+ mice partially mimic the human phenotype and therefore represent a relevant model for further investigations of the physiopathology of cortical malformations.
- MeSH
- centrozom metabolismus MeSH
- chování zvířat MeSH
- elektronová mikroskopie MeSH
- embryo savčí MeSH
- epilepsie genetika MeSH
- fibroblasty cytologie metabolismus ultrastruktura MeSH
- genetická predispozice k nemoci MeSH
- genový knockin MeSH
- HeLa buňky MeSH
- intravitální mikroskopie MeSH
- konfokální mikroskopie MeSH
- lidé MeSH
- malformace mozkové kůry genetika MeSH
- mikrotubuly genetika metabolismus MeSH
- missense mutace MeSH
- modely nemocí na zvířatech MeSH
- mozková kůra abnormality cytologie diagnostické zobrazování MeSH
- myši transgenní MeSH
- myši MeSH
- neurogeneze genetika MeSH
- neurony fyziologie MeSH
- pohyb buněk genetika MeSH
- tubulin genetika MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- audiovizuální média MeSH
- časopisecké články MeSH
- práce podpořená grantem MeSH
Colletotrichum species are known as important pathogens of plants with an impact on crop production. Some of these species are also known as a cause of rare ophthalmic infections in humans. A case of keratitis caused by Colletotrichum dematium after corneal trauma in a 56-year-old woman is presented. Infection was diagnosed based on positive microscopy and culture. The fungal isolate was identified by morphological characteristics and DNA sequencing of the ITS rDNA region, β-tubulin (tub2) and glyceraldehyde-3-phosphate dehydrogenase (gapdh) genes. The patient responded well to topical therapy with amphotericin B combined with intravenous amphotericin B but improvement was associated with the corneal collagen cross-linking. The review of the literature revealed another 13 cases of C. dematium keratitis, all but one patient having at least one keratitis risk factor in their history. Almost all patients (n = 12) were treated with topical polyene antibiotics (natamycin or amphotericin B), improvement and cure were achieved in eight of them.
- MeSH
- amfotericin B aplikace a dávkování MeSH
- antifungální látky aplikace a dávkování MeSH
- aplikace lokální MeSH
- Colletotrichum klasifikace genetika izolace a purifikace MeSH
- diagnostické techniky molekulární MeSH
- dospělí MeSH
- glyceraldehyd-3-fosfátdehydrogenasa (fosforylační) genetika MeSH
- keratitida diagnóza mikrobiologie patologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- mezerníky ribozomální DNA genetika MeSH
- mikrobiologické techniky MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mykózy diagnóza mikrobiologie patologie MeSH
- poranění oka komplikace MeSH
- sekvenční analýza DNA MeSH
- tubulin genetika MeSH
- výsledek terapie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
- systematický přehled MeSH
Posttranslational modifications of tubulin are emerging regulators of microtubule functions. We have shown earlier that upregulated polyglutamylation is linked to rapid degeneration of Purkinje cells in mice with a mutation in the deglutamylating enzyme CCP1. How polyglutamylation leads to degeneration, whether it affects multiple neuron types, or which physiological processes it regulates in healthy neurons has remained unknown. Here, we demonstrate that excessive polyglutamylation induces neurodegeneration in a cell-autonomous manner and can occur in many parts of the central nervous system. Degeneration of selected neurons in CCP1-deficient mice can be fully rescued by simultaneous knockout of the counteracting polyglutamylase TTLL1. Excessive polyglutamylation reduces the efficiency of neuronal transport in cultured hippocampal neurons, suggesting that impaired cargo transport plays an important role in the observed degenerative phenotypes. We thus establish polyglutamylation as a cell-autonomous mechanism for neurodegeneration that might be therapeutically accessible through manipulation of the enzymes that control this posttranslational modification.
- MeSH
- aktivní transport genetika MeSH
- myši knockoutované MeSH
- myši MeSH
- neurodegenerativní nemoci genetika metabolismus patologie MeSH
- peptidsynthasy genetika metabolismus MeSH
- peptidy genetika metabolismus MeSH
- posttranslační úpravy proteinů * MeSH
- proteiny nervové tkáně genetika metabolismus MeSH
- Purkyňovy buňky metabolismus patologie MeSH
- tubulin genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Invasive fungal disease represents one of the severe complications in haematopoietic stem cell transplant recipients. We describe a case of a patient treated for relapse of chronic lymphoblastic leukaemia 6 years after HSCT. The patient was treated for invasive pulmonary aspergillosis but died 3 months later from multiple organ failures consisting of haemorrhagic necrotizing fungal pneumonia, refractory chronic hepatic graft versus host disease and cytomegalovirus hepatitis. Autopsy samples revealed histopathological evidence of fungal hyphae and an unusual Aspergillus nidulans-like species was isolated in pure culture. More precise identification was achieved by using scanning electron microscopy of ascospores and sequencing of calmodulin gene, and the isolate was subsequently re-identified as A. sublatus (section Nidulantes) and showed good in vitro susceptibility against all classes of antifungals. Commonly used ITS rDNA region and β-tubulin gene fail to discriminate A. sublatus from related pathogenic species, especially A. quadrilineatus and A. nidulans. Although this is the first case of proven IPA attributed to A. sublatus, we demonstrated that at least some previously reported infections due to A. quadrilineatus were probably caused by this cryptic species.
- MeSH
- antifungální látky aplikace a dávkování MeSH
- Aspergillus klasifikace cytologie genetika izolace a purifikace MeSH
- chronická lymfatická leukemie komplikace MeSH
- cytomegalovirové infekce komplikace diagnóza MeSH
- DNA fungální chemie genetika MeSH
- fatální výsledek MeSH
- fylogeneze MeSH
- invazivní plicní aspergilóza diagnóza farmakoterapie mikrobiologie patologie MeSH
- kalmodulin genetika MeSH
- lidé středního věku MeSH
- lidé MeSH
- mezerníky ribozomální DNA chemie genetika MeSH
- mikrobiální testy citlivosti MeSH
- mikroskopie elektronová rastrovací MeSH
- nemoc štěpu proti hostiteli komplikace diagnóza MeSH
- příjemce transplantátu MeSH
- sekvenční analýza DNA MeSH
- shluková analýza MeSH
- transplantace hematopoetických kmenových buněk škodlivé účinky MeSH
- tubulin genetika MeSH
- virová hepatitida u lidí komplikace diagnóza MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
γ-Tubulin is essential for microtubule nucleation and also plays less understood roles in nuclear and cell-cycle-related functions. High abundancy of γ-tubulin in acentrosomal Arabidopsis cells facilitated purification and biochemical characterization of large molecular species of γ-tubulin. TEM, fluorescence, and atomic force microscopy of purified high molecular γ-tubulin forms revealed the presence of linear filaments with a double protofilament substructure, filament bundles and aggregates. Filament formation from highly purified γ-tubulin free of γ-tubulin complex proteins (GCPs) was demonstrated for both plant and human γ-tubulin. Moreover, γ-tubulin associated with porcine brain microtubules formed oligomers. Experimental evidence on the intrinsic ability of γ-tubulin to oligomerize/polymerize was supported by conservation of α- and β-tubulin interfaces for longitudinal and lateral interactions for γ-tubulins. STED (stimulated emission depletion) microscopy of Arabidopsis cells revealed fine, short γ-tubulin fibrillar structures enriched on mitotic microtubular arrays that accumulated at polar regions of acentrosomal spindles and the outer nuclear envelope before mitosis, and were also present in nuclei. Fine fibrillar structures of γ-tubulin representing assemblies of higher order were localized in cell-cycle-dependent manner at sites of dispersed γ-tubulin location in acentrosomal plant cells as well as at sites of local γ-tubulin enrichment after drug treatment. Our findings that γ-tubulin preserves the capability of prokaryotic tubulins to self-organize into filaments assembling by lateral interaction into bundles/clusters help understanding of the relationship between structure and multiple cellular functions of this protein species and suggest that besides microtubule nucleation and organization, γ-tubulin may also have scaffolding or sequestration functions.
- MeSH
- Arabidopsis chemie genetika MeSH
- cytoskelet chemie genetika MeSH
- mikrofilamenta chemie genetika ultrastruktura MeSH
- mikrotubuly chemie genetika MeSH
- mitóza genetika MeSH
- polymerizace MeSH
- proteinové agregáty genetika MeSH
- proteiny asociované s mikrotubuly chemie genetika MeSH
- tubulin chemie genetika ultrastruktura MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH