Osteoblasts orchestrate bone formation through the secretion of type I collagen and other constituents of the matrix on which hydroxyapatite crystals mineralize. Here, we show that TENT5A, whose mutations were found in congenital bone disease osteogenesis imperfecta patients, is a cytoplasmic poly(A) polymerase playing a crucial role in regulating bone mineralization. Direct RNA sequencing revealed that TENT5A is induced during osteoblast differentiation and polyadenylates mRNAs encoding Col1α1, Col1α2, and other secreted proteins involved in osteogenesis, increasing their expression. We postulate that TENT5A, possibly together with its paralog TENT5C, is responsible for the wave of cytoplasmic polyadenylation of mRNAs encoding secreted proteins occurring during bone mineralization. Importantly, the Tent5a knockout (KO) mouse line displays bone fragility and skeletal hypomineralization phenotype resulting from quantitative and qualitative collagen defects. Thus, we report a biologically relevant posttranscriptional regulator of collagen production and, more generally, bone formation.

• MeSH
• buněčná diferenciace MeSH
• fyziologická kalcifikace genetika   MeSH
• kolagen typu I, řetězec alfa 1 genetika   metabolismus   MeSH
• kolagen typu I genetika   metabolismus   MeSH
• lidé MeSH
• messenger RNA genetika   metabolismus   MeSH
• modely nemocí na zvířatech MeSH
• myši knockoutované MeSH
• myši MeSH
• neurotrofní faktory genetika   metabolismus   MeSH
• nukleotidyltransferasy genetika   metabolismus   MeSH
• oční proteiny genetika   metabolismus   MeSH
• osteoblasty metabolismus   patologie   MeSH
• osteogenesis imperfecta genetika   metabolismus   patologie   MeSH
• osteogeneze genetika   MeSH
• osteonektin genetika   metabolismus   MeSH
• poly(A)-polymerasa genetika   metabolismus   MeSH
• polyadenylace MeSH
• protein - isoformy nedostatek   genetika   MeSH
• sekvenční analýza RNA MeSH
• serpiny genetika   metabolismus   MeSH
• signální transdukce MeSH
• vývojová regulace genové exprese MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• chronická nemoc MeSH
• diabetes mellitus 1. typu komplikace   MeSH
• exprese genu MeSH
• finanční podpora výzkumu jako téma MeSH
• kardiomyopatie MeSH
• krysa rodu rattus MeSH
• messenger RNA genetika   MeSH
• modely nemocí na zvířatech MeSH
• neuropeptid Y fyziologie   MeSH
• receptory neuropeptidů MeSH
• srdce MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH

Termination codons in mRNA molecules are typically specified directly by the sequence of the corresponding gene. However, in mitochondria of a few eukaryotic groups, some mRNAs contain the termination codon UAA deriving one or both adenosines from transcript polyadenylation. Here, we show that a similar phenomenon occurs for a substantial number of nuclear genes in Blastocystis spp., divergent unicellular eukaryote gut parasites. Our analyses of published genomic data from Blastocystis sp. subtype 7 revealed that polyadenylation-mediated creation of termination codons occurs in approximately 15% of all nuclear genes. As this phenomenon has not been noticed before, the procedure previously employed to annotate the Blastocystis nuclear genome sequence failed to correctly define the structure of the 3'-ends of hundreds of genes. From sequence data we have obtained from the distantly related Blastocystis sp. subtype 1 strain, we show that this phenomenon is widespread within the Blastocystis genus. Polyadenylation in Blastocystis appears to be directed by a conserved GU-rich element located four nucleotides downstream of the polyadenylation site. Thus, the highly precise positioning of the polyadenylation in Blastocystis has allowed reduction of the 3'-untranslated regions to the point that, in many genes, only one or two nucleotides of the termination codon are left.

• MeSH
• Blastocystis chemie   genetika   MeSH
• blastocystóza parazitologie   MeSH
• lidé MeSH
• messenger RNA chemie   genetika   MeSH
• molekulární sekvence - údaje MeSH
• polyadenylace * MeSH
• protozoální proteiny chemie   genetika   MeSH
• sekvence aminokyselin MeSH
• sekvence nukleotidů MeSH
• terminační kodon chemie   genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• dopisy MeSH
• práce podpořená grantem MeSH

Alternative polyadenylation is an important and pervasive mechanism that generates heterogeneous 3'-termini of mRNA and is considered an important regulator of gene expression. We performed bioinformatics analyses of ESTs and the 3'-UTRs of the main transcript splice variants of the translational initiation factor eIF4E1 and its family members, eIF4E2 and eIF4E3. This systematic analysis led to the prediction of new polyadenylation signals. All identified polyadenylation sites were subsequently verified by 3'RACE of transcripts isolated from human lymphoblastic cell lines. This led to the observation that multiple simultaneous polyadenylation site utilization occurs in single cell population. Importantly, we described the use of new polyadenylation site in the eIF4E1 mRNA, which lacked any known polyadenylation signal. The proportion of eIF4E1 transcripts derived from the first two polyadenylation sites in eIF4E1 mRNA achieved 15% in a wide range of cell lines. This result demonstrates the ubiquitous presence of ARE-lacking transcripts, which escape HuR/Auf1-mediated control, the main mechanism of eIF4E1 gene expression regulation. We found many EST clones documenting the significant production of transcript variants 2-4 of eIF4E2 gene that encode proteins with C-termini that were distinct from the mainly studied prototypical isoform A. Similarly, eIF4E3 mRNAs are produced as two main variants with the same very long 3'-UTR with potential for heavy post-transcriptional regulation. We identified sparsely documented transcript variant 1 of eIF4E3 gene in human placenta. eIF4E3 truncated transcript variants were found mainly in brain. We propose to elucidate the minor splice variants of eIF4E2 and eIF4E3 in great detail because they might produce proteins with modified features that fulfill different cellular roles from their major counterparts.

• MeSH
• 3' nepřekládaná oblast MeSH
• buněčné linie MeSH
• eukaryotický iniciační faktor 4E genetika   metabolismus   MeSH
• exprimované sekvenční adresy MeSH
• lidé MeSH
• messenger RNA genetika   metabolismus   MeSH
• mozek metabolismus   MeSH
• placenta metabolismus   MeSH
• polyadenylace genetika   MeSH
• proteiny vázající čepičku mRNA genetika   MeSH
• regulace genové exprese MeSH
• sestřih RNA genetika   MeSH
• těhotenství MeSH
• Check Tag
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Strigolactones (SL) contribute to drought acclimatization in shoots, because SL-depleted plants are hypersensitive to drought due to stomatal hyposensitivity to abscisic acid (ABA). However, under drought, SL biosynthesis is repressed in roots, suggesting organ specificity in their metabolism and role. Because SL can be transported acropetally, such a drop may also affect shoots, as a systemic indication of stress. We investigated this hypothesis by analysing molecularly and physiologically wild-type (WT) tomato (Solanum lycopersicum) scions grafted onto SL-depleted rootstocks, compared with self-grafted WT and SL-depleted genotypes, during a drought time-course. Shoots receiving few SL from the roots behaved as if under mild stress even if irrigated. Their stomata were hypersensitive to ABA (likely via a localized enhancement of SL synthesis in shoots). Exogenous SL also enhanced stomata sensitivity to ABA. As the partial shift of SL synthesis from roots to shoots mimics what happens under drought, a reduction of root-produced SL might represent a systemic signal unlinked from shootward ABA translocation, and sufficient to prime the plant for better stress avoidance.

• MeSH
• biologické modely MeSH
• biosyntetické dráhy genetika   MeSH
• dehydratace MeSH
• fenotyp MeSH
• fyziologický stres * genetika   MeSH
• kořeny rostlin metabolismus   MeSH
• kyselina abscisová metabolismus   MeSH
• laktony metabolismus   MeSH
• listy rostlin fyziologie   MeSH
• messenger RNA genetika   metabolismus   MeSH
• období sucha * MeSH
• regulace genové exprese u rostlin MeSH
• rostlinné geny MeSH
• signální transdukce * MeSH
• Solanum lycopersicum genetika   fyziologie   MeSH
• transpirace rostlin MeSH
• voda fyziologie   MeSH
• výhonky rostlin genetika   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH

Although the involvement of the extracellular signal-regulated kinases 1 and 2 (ERK1/2) pathway in the regulation of cytostatic factor (CSF) activity; as well as in microtubules organization during meiotic maturation of oocytes; has already been described in detail; rather less attention has been paid to the role of ERK1/2 in the regulation of mRNA translation. However; important data on the role of ERK1/2 in translation during oocyte meiosis have been documented. This review focuses on recent findings regarding the regulation of translation and the role of ERK1/2 in this process in the meiotic cycle of mammalian oocytes. The specific role of ERK1/2 in the regulation of mammalian target of rapamycin (mTOR); eukaryotic translation initiation factor 4E (eIF4E) and cytoplasmic polyadenylation element binding protein 1 (CPEB1) activity is addressed along with additional focus on the other key players involved in protein translation.

• MeSH
• cytoplazma genetika   metabolismus   MeSH
• eukaryotický iniciační faktor 4E metabolismus   MeSH
• faktory štěpení a polyadenylace mRNA metabolismus   MeSH
• fosfatidylinositol-3-kinasy metabolismus   MeSH
• lidé MeSH
• meióza * MeSH
• messenger RNA genetika   metabolismus   MeSH
• mitogenem aktivovaná proteinkinasa 1 metabolismus   MeSH
• mitogenem aktivovaná proteinkinasa 3 metabolismus   MeSH
• mitogenem aktivované proteinkinasy metabolismus   MeSH
• oocyty metabolismus   MeSH
• polyadenylace MeSH
• proteosyntéza * MeSH
• signální transdukce MeSH
• TOR serin-threoninkinasy metabolismus   MeSH
• vazba proteinů MeSH
• vývojová regulace genové exprese MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Cerebral ischemia-reperfusion injury (CIRI) is the predominant cause of neurological disability after cardiac arrest/cardiopulmonary resuscitation (CA/CPR). The endoplasmic reticulum stress (ERs)-induced apoptosis plays an important role in neuronal survival/death in CIRI. Our previous studies reported that the extracellular signal-regulated kinase (ERK) inhibitor, PD98059, alleviates CIRI after CA/CPR. Whether ERs-induced apoptosis is involved in the neuroprotection of PD98059 remains unknown. This study aims to investigate the effects of ERK inhibition by PD98059 on ERs-induced apoptosis after CIRI in the CA/CPR rat model. The baseline characteristics of male adult Sprague-Dawley (SD) rats in all groups were evaluated before CA/CPR. The SD rats that survived from CA/CPR were randomly divided into 3 groups (n=12/group): normal saline group (1 ml/kg), dimethylsulfoxide (DMSO, the solvent of PD98059, 1 ml/kg) group, PD98059 group (0.3 mg/kg). Another 12 SD rats were randomly selected as the Sham group. Twenty-four hours after resuscitation, neural injury was assessed by survival rate, neurological deficit scores (NDS) and Nissl staining; apoptosis of brain cells was detected using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining; mRNA expression and protein levels of ERs-related protein BIP, PERK, ATF4 and CHOP were checked with RT-PCR and Western Blot. The results showed that there were no significant differences in baseline characteristics before CA/CPR among all groups. PD98059 significantly improved survival rate and NDS, increased the Nissl bodies in neurons, reduced apoptosis, downregulated the mRNA transcription and expression levels of BIP, PERK, ATF4 and CHOP at 24 h after CA/CPR. Our results demonstrate that inhibition of ERK by PD98059 alleviates ERs-induced apoptosis via BIP-PERK-ATF4-CHOP signaling pathway and mitigates CIRI in the CA/CPR rat model.

• MeSH
• apoptóza MeSH
• extracelulárním signálem regulované MAP kinasy MeSH
• krysa rodu rattus MeSH
• messenger RNA MeSH
• poranění mozku * MeSH
• potkani Sprague-Dawley MeSH
• reperfuzní poškození * metabolismus   MeSH
• srdeční zástava * komplikace   MeSH
• stres endoplazmatického retikula MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

There is a growing body of evidence that cytokines contribute to both induction and maintenance of neuropathic pain derived from changes in dorsal root ganglia (DRG), including the activity of the primary sensory neurons and their satellite glial cells (SGC). We used immunofluorescence and in situ hybridization methods to provide evidence that chronic constriction injury (CCI) of the sciatic nerve induces synthesis of interleukin-6 (IL-6) in SGC, elevation of IL-6 receptor (IL-6R) and activation of signal transducer and activator of transcription 3 (STAT3) signalling. Unilateral CCI of the rat sciatic nerve induced mechanoallodynia and thermal hyperalgesia in ipsilateral hind paws, but contralateral paws exhibited only temporal changes of sensitivity. We demonstrated that IL-6 mRNA and protein, which were expressed at very low levels in naïve DRG, were bilaterally increased not only in L4-L5 DRG neurons but also in SGC activated by unilateral CCI. Besides IL-6, substantial increase of IL-6R and pSTAT3 expression occurred in SGC following CCI, however, IL-6R associated protein, gp130 levels did not change. The results may suggest that unilateral CCI of the sciatic nerve induces bilateral activation of SGC in L4-L5 DRG to transduce IL-6 signalling during neuroinflammation.

• MeSH
• cytokinový receptor gp130 genetika   metabolismus   MeSH
• funkční lateralita MeSH
• interleukin-6 genetika   metabolismus   MeSH
• krysa rodu rattus MeSH
• měření bolesti MeSH
• messenger RNA metabolismus   MeSH
• modely nemocí na zvířatech MeSH
• nemoci sedacího nervu patologie   patofyziologie   MeSH
• neuroglie metabolismus   MeSH
• potkani Wistar MeSH
• receptory interleukinu-6 genetika   metabolismus   MeSH
• regulace genové exprese fyziologie   MeSH
• signální transdukce fyziologie   MeSH
• spinální ganglia patologie   patofyziologie   MeSH
• transkripční faktor STAT3 metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Non-coding RNA polymerase II transcripts are processed by the poly(A)-independent termination pathway that requires the Nrd1 complex. The Nrd1 complex includes two RNA-binding proteins, the nuclear polyadenylated RNA-binding (Nab) 3 and the nuclear pre-mRNA down-regulation (Nrd) 1 that bind their specific termination elements. Here we report the solution structure of the RNA-recognition motif (RRM) of Nab3 in complex with a UCUU oligonucleotide, representing the Nab3 termination element. The structure shows that the first three nucleotides of UCUU are accommodated on the β-sheet surface of Nab3 RRM, but reveals a sequence-specific recognition only for the central cytidine and uridine. The specific contacts we identified are important for binding affinity in vitro as well as for yeast viability. Furthermore, we show that both RNA-binding motifs of Nab3 and Nrd1 alone bind their termination elements with a weak affinity. Interestingly, when Nab3 and Nrd1 form a heterodimer, the affinity to RNA is significantly increased due to the cooperative binding. These findings are in accordance with the model of their function in the poly(A) independent termination, in which binding to the combined and/or repetitive termination elements elicits efficient termination.

• MeSH
• genetická transkripce MeSH
• jaderné proteiny chemie   genetika   metabolismus   MeSH
• konformace proteinů MeSH
• magnetická rezonanční spektroskopie MeSH
• multimerizace proteinu MeSH
• oligonukleotidy chemie   metabolismus   MeSH
• proteiny vázající RNA chemie   genetika   metabolismus   MeSH
• roztoky MeSH
• Saccharomyces cerevisiae - proteiny chemie   genetika   metabolismus   MeSH
• Saccharomyces cerevisiae genetika   MeSH
• sekvence nukleotidů MeSH
• vazba proteinů MeSH
• vazebná místa MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The activity of the Wnt pathway undergoes complex regulation to ensure proper functioning of this principal signaling mechanism during development of adult tissues. The regulation may occur at several levels and includes both positive and negative feedback loops. In the present study we employed one of such negative feedback regulators, naked cuticle homolog 1 (Nkd1), to follow the Wnt pathway activity in the intestine and liver and in neoplasia originated in these organs. Using lineage tracing in transgenic mice we localized Nkd1 mRNA to the bottom parts of the small intestinal crypts and hepatocytes surrounding the central vein of the hepatic lobule. Furthermore, in two mouse models of intestinal tumorigenesis, Nkd1 expression levels were elevated in tumors when compared to healthy tissue. We utilized a collection of human intestinal polyps and carcinomas to confirm that NKD1 represents a robust marker of neoplastic growth. In addition, expression analysis of NKD1 in liver cancer showed that high expression levels of the gene distinguish a subclass of hepatocellular carcinomas related to aberrant Wnt signaling. Finally, our results were confirmed by bioinformatic analysis of large publicly available datasets that included gene expression profiling and high-throughput sequencing data of human colon and liver cancer specimens.

• MeSH
• beta-katenin genetika   metabolismus   MeSH
• genetická transkripce MeSH
• hepatocelulární karcinom metabolismus   patologie   MeSH
• hepatocyty metabolismus   patologie   MeSH
• kolorektální nádory metabolismus   patologie   MeSH
• lidé MeSH
• messenger RNA metabolismus   MeSH
• mutace MeSH
• myši transgenní MeSH
• myši MeSH
• nádory jater metabolismus   patologie   MeSH
• protein familiární adenomatózní polypózy nedostatek   genetika   metabolismus   MeSH
• proteiny Wnt metabolismus   MeSH
• signální transdukce MeSH
• střevní nádory metabolismus   patologie   MeSH
• transportní proteiny genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH