Vyhodnocení telomerázové aktivity a měření telomer by mohlo patřit k novým perspektivním prognostickým faktorům u mnohočetného myelomu. Možnost využít hTERT jako nádorového antigenu by výrazným způsobem obohatila vakcinační strategii u mnohočetného myelomu, kde je doposud limitací nízká účinnost, respektive nepřítomnost silného antigenu.; Telomerase assay in multiple myeloma is new perspective prognostic factor in multiple myeloma The use od hTERT as tumor antigen is promising new treatment modality and could circumvent limitation of current vaccination protocols in multiple myeloma in which low immunogenic antigens and consequently minimal responses are reported.

• MeSH
• mnohočetný myelom MeSH
• prognóza MeSH
• telomerasa MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• alergologie a imunologie
• onkologie
• hematologie a transfuzní lékařství
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR

Telomeres, essential for maintaining genomic stability, are typically preserved through the action of telomerase, a ribonucleoprotein complex that synthesizes telomeric DNA. One of its two core components, telomerase RNA (TR), serves as the template for this synthesis, and its evolution across different species is both complex and diverse. This review discusses recent advancements in understanding TR evolution, with a focus on plants (Viridiplantae). Utilizing novel bioinformatic tools and accumulating genomic and transcriptomic data, combined with corresponding experimental validation, researchers have begun to unravel the intricate pathways of TR evolution and telomere maintenance mechanisms. Contrary to previous beliefs, a monophyletic origin of TR has been demonstrated first in land plants and subsequently across the broader phylogenetic megagroup Diaphoretickes. Conversely, the discovery of plant-type TRs in insects challenges assumptions about the monophyletic origin of TRs in animals, suggesting evolutionary innovations coinciding with arthropod divergence. The review also highlights key challenges in TR identification and provides examples of how these have been addressed. Overall, this work underscores the importance of expanding beyond model organisms to comprehend the full complexity of telomerase evolution, with potential applications in agriculture and biotechnology.

• MeSH
• fylogeneze MeSH
• molekulární evoluce * MeSH
• RNA * genetika   metabolismus   MeSH
• rostliny genetika   MeSH
• telomerasa * genetika   metabolismus   MeSH
• telomery * metabolismus   genetika   MeSH
• Viridiplantae genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

In contrast to the catalytic subunit of telomerase, its RNA subunit (TR) is highly divergent in size, sequence and biogenesis pathways across eukaryotes. Current views on TR evolution assume a common origin of TRs transcribed with RNA polymerase II in Opisthokonta (the supergroup including Animalia and Fungi) and Trypanosomida on one hand, and TRs transcribed with RNA polymerase III under the control of type 3 promoter, found in TSAR and Archaeplastida supergroups (including e.g. ciliates and Viridiplantae taxa, respectively). Here, we focus on unknown TRs in one of the largest Animalia order - Hymenoptera (Arthropoda) with more than 300 available representative genomes. Using a combination of bioinformatic and experimental approaches, we identify their TRs. In contrast to the presumed type of TRs (H/ACA box snoRNAs transcribed with RNA Polymerase II) corresponding to their phylogenetic position, we find here short TRs of the snRNA type, likely transcribed with RNA polymerase III under the control of the type 3 promoter. The newly described insect TRs thus question the hitherto assumed monophyletic origin of TRs across Animalia and point to an evolutionary switch in TR type and biogenesis that was associated with the divergence of Arthropods.

• MeSH
• Eukaryota genetika   MeSH
• fylogeneze MeSH
• Hymenoptera * genetika   MeSH
• konformace nukleové kyseliny MeSH
• RNA-polymerasa II genetika   metabolismus   MeSH
• RNA-polymerasa III genetika   metabolismus   MeSH
• RNA genetika   MeSH
• rostliny genetika   MeSH
• telomerasa * genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH

BACKGROUND: Mutations in the LDLR gene are the most frequent cause of Familial hypercholesterolemia, an autosomal dominant disease characterised by elevated concentrations of LDL in blood plasma. In many populations, large genomic rearrangements account for approximately 10% of mutations in the LDLR gene. METHODS: DNA diagnostics of large genomic rearrangements was based on Multiple Ligation dependent Probe Amplification (MLPA). Subsequent analyses of deletion and duplication breakpoints were performed using long-range PCR, PCR, and DNA sequencing. RESULTS: In set of 1441 unrelated FH patients, large genomic rearrangements were found in 37 probands. Eight different types of rearrangements were detected, from them 6 types were novel, not described so far. In all rearrangements, we characterized their exact extent and breakpoint sequences. CONCLUSIONS: Sequence analysis of deletion and duplication breakpoints indicates that intrachromatid non-allelic homologous recombination (NAHR) between Alu elements is involved in 6 events, while a non-homologous end joining (NHEJ) is implicated in 2 rearrangements. Our study thus describes for the first time NHEJ as a mechanism involved in genomic rearrangements in the LDLR gene.

• MeSH
• elementy Alu MeSH
• genová přestavba MeSH
• hyperlipoproteinemie typ II genetika   MeSH
• LDL-receptory genetika   MeSH
• lidé MeSH
• molekulární sekvence - údaje MeSH
• sekvence nukleotidů MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

• MeSH
• lékařská genetika MeSH
• lidé MeSH
• Nobelova cena MeSH
• replikace DNA genetika   MeSH
• ribozomální DNA MeSH
• stárnutí buněk genetika   MeSH
• telomerasa genetika   MeSH
• telomery genetika   MeSH
• Check Tag
• lidé MeSH

BACKGROUND: Epidermolysis bullosa simplex (EBS) is an inherited skin disorder caused by mutations in the keratin 5 (KRT5) and keratin 14 (KRT14) genes, with fragility of basal keratinocytes leading to epidermal cytolysis and blistering. OBJECTIVES: In this study, we characterized mutations in KRT5 and KRT14 genes in patients with EBS and investigated their possible structure-function correlations. MATERIALS AND METHODS: Mutations were characterized using polymerase chain reaction (PCR) and DNA sequencing. Further, to explore possible correlations with function, the structural effects of the mutations in segment 2B of KRT5 and KRT14 and associated with EBS in our patients, as well as those reported previously, were modelled by molecular dynamics with the aid of the known crystal structure of the analogous segment of human vimentin. RESULTS: We have identified mutations in the KRT5 and KRT14 genes in 16 of 23 families affected by EBS in the Czech Republic. Eleven different sequence variants were found, of which four have not been reported previously. Novel mutations were found in two patients with the EBS-Dowling-Meara variant (EBS-DM) [KRT14-p.Ser128Pro and KRT14-p.Gln374_Leu387dup(14)] and in three patients with localized EBS (KRT14-p.Leu136Pro and KRT5-p.Val143Ala). Molecular dynamics studies show that the mutations p.Glu411del and p.Ile467Thr perturb the secondary alpha-helical structure of the mutated polypeptide chain, the deletion p.Glu411del in KRT14 has a strong but only local influence on the secondary structure of KRT14, and the structural impact of the mutation p.Ile467Thr in KRT5 is spread along the helix to the C-terminus. In all the other point mutations studied, the direct structural impact was significantly weaker and did not destroy the alpha-helical pattern of the secondary protein structure. The changes of 3-D structure of the KRT5/KRT14 dimer induced by the steric structural impact of the single point mutations, and the resulting altered inter- and intramolecular contacts, are spread along the protein helices to the protein C-terminus, but the overall alpha-helical character of the secondary structure is not destroyed and the atomic displacements induced by mutations cause only limited-scale changes of the quaternary structure of the dimer. CONCLUSIONS: The results of molecular modelling show relationships between patients' phenotypes and the structural effects of individual mutations.

• MeSH
• dítě MeSH
• dospělí MeSH
• epidermolysis bullosa simplex genetika   patologie   MeSH
• fenotyp MeSH
• fluorescenční mikroskopie MeSH
• genetická predispozice k nemoci MeSH
• intermediární filamenta ultrastruktura   MeSH
• keratin-14 genetika   MeSH
• keratin-5 genetika   MeSH
• kůže ultrastruktura   MeSH
• lidé MeSH
• molekulární modely MeSH
• mutace MeSH
• předškolní dítě MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The TERT (telomerase reverse transcriptase) subunit of telomerase is an intensively studied macromolecule due to its key importance in maintaining genome integrity and role in cellular aging and cancer. In an effort to provide an up-to-date overview of the topic, we discuss the structure of TERT genes, their alternative splicing products and their functions. Nucleotide databases contain more than 90 full-length cDNA sequences of telomerase protein subunits. Numerous in silico, in vitro and in vivo experimental techniques have revealed a great deal of structural and functional data describing particular features of the telomerase subunit in various model organisms. We explore whether particular findings are generally applicable to telomerases or species-specific. We also discuss in an evolutionary context the role of identified functional TERT subdomains.

• MeSH
• alternativní sestřih MeSH
• eukaryotické buňky enzymologie   chemie   MeSH
• lidé MeSH
• molekulární evoluce MeSH
• molekulární sekvence - údaje MeSH
• prokaryotické buňky enzymologie   chemie   MeSH
• telomerasa genetika   chemie   metabolismus   MeSH
• telomery metabolismus   MeSH
• vazba proteinů MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH

BACKGROUND: Telomerase activity is associated with many malignancies, including head and neck cancer. The use of telomerase activity as a diagnostic and prognostic marker of head and neck cancer development was examined and compared with standard histological analysis. PATIENTS AND METHODS: Telomerase activity was determined using quantitative dual-colour real-time TRAP (telomeric repeat amplification protocol). In each of 58 patients, a sample of tumour tissue, adjacent mucosa and normal muscle was collected. RESULTS: Telomerase activation was observed in 88% of tumour tissues and 34% of tumour-adjacent mucosa samples. No telomerase activity was detected in normal muscle tissues. Telomerase activity correlated with tumour grade, with an average of 4.6 telomerase units (T.U.) in well-differentiated, 8.3 T.U. in moderately-differentiated and 20 T.U. in poorly differentiated tumours. Relapse occurred in 13 patients and no telomerase activity was detected in 3 recurrent tumours. CONCLUSION: Telomerase activity may be used as an objective parameter inversely related to tumour differentiation. Prognosis in telomerase-negative tumours is worse than that of the telomerase-positive group.

• MeSH
• kouření genetika   metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádorové biomarkery metabolismus   MeSH
• nádory hlavy a krku enzymologie   genetika   MeSH
• prognóza MeSH
• senioři MeSH
• telomerasa metabolismus   MeSH
• telomery metabolismus   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• práce podpořená grantem MeSH

Telomeres are nucleoprotein structures ensuring the stability of eukaryotic chromosome ends. Two protein families, TRFL (TFL-Like) and SMH (Single-Myb-Histone), containing a specific telobox motif in their Myb domain, have been identified as potential candidates involved in a functional nucleoprotein structure analogous to human "shelterin" at plant telomeres. We analyze the DNA-protein interaction of the full-length and truncated variants of AtTRB1, a SMH-family member with a typical structure: N-terminal Myb domain, central H1/5 domain and C-terminal coiled-coil. We show that preferential interaction of AtTRB1 with double-stranded telomeric DNA is mediated by the Myb domain, while the H1/5 domain is involved in non-specific DNA-protein interaction and in the multimerization of AtTRB1.

• MeSH
• Arabidopsis genetika   metabolismus   MeSH
• DNA rostlinná metabolismus   MeSH
• DNA vazebné proteiny genetika   metabolismus   MeSH
• elektroforéza v polyakrylamidovém gelu MeSH
• financování organizované MeSH
• histony metabolismus   MeSH
• klonování DNA MeSH
• lidé MeSH
• onkogenní proteiny v-myb metabolismus   MeSH
• proteiny huseníčku genetika   metabolismus   MeSH
• telomery metabolismus   MeSH
• vazba proteinů MeSH
• Check Tag
• lidé MeSH

Limb girdle muscular dystrophy type 2A (LGMD2A) is caused by single or small nucleotide changes widespread along the CAPN3 gene, which encodes the muscle-specific proteolytic enzyme calpain-3. About 356 unique allelic variants of CAPN3 have been identified to date. We performed analysis of the CAPN3 gene in LGMD2A patients at both the mRNA level using reverse transcription-PCR, and at the DNA level using PCR and denaturing high performance liquid chromatography. In four patients, we detected homozygous occurrence of a missense mutation or an in-frame deletion at the mRNA level although the DNA was heterozygous for this mutation in conjunction with a frame-shift mutation. The relationship observed in 12 patients between the quantity of CAPN3 mRNA, determined using real-time PCR, and the genotype leads us to propose that CAPN3 mRNAs which contain frame-shift mutations are degraded by nonsense-mediated mRNA decay. Our results illustrate the importance of DNA analysis for reliable establishment of mutation status, and provide a new insight into the process of mRNA decay in cells of LGMD2A patients.

• MeSH
• delece genu MeSH
• dítě MeSH
• DNA biosyntéza   genetika   MeSH
• dospělí MeSH
• financování organizované MeSH
• genotyp MeSH
• introny genetika   MeSH
• kalpain genetika   MeSH
• kojenec MeSH
• lidé MeSH
• messenger RNA genetika   MeSH
• mladiství MeSH
• nesmyslný kodon genetika   MeSH
• pletencové svalové dystrofie genetika   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• posunová mutace genetika   MeSH
• předškolní dítě MeSH
• svalové proteiny genetika   MeSH
• western blotting MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH