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
The interleukin 17 (IL-17) cytokine and receptor family is central to antimicrobial resistance and inflammation in the lung. Mice lacking IL-17A, IL-17F, or the IL-17RA subunit were compared with wild-type mice for susceptibility to airway inflammation in models of infection and allergy. Signaling through IL-17RA was required for efficient microbial clearance and prevention of allergy; in the absence of IL-17RA, signaling through IL-17RC on epithelial cells, predominantly by IL-17F, significantly exacerbated lower airway Aspergillus or Pseudomonas infection and allergic airway inflammation. In contrast, following infection with the upper respiratory pathogen Staphylococcus aureus, the IL-17F/IL-17RC axis mediated protection. Thus, IL-17A and IL-17F exert distinct biological effects during pulmonary infection; the IL-17F/IL-17RC signaling axis has the potential to significantly worsen pathogen-associated inflammation of the lower respiratory tract in particular, and should be investigated further as a therapeutic target for treating pathological inflammation in the lung.
- MeSH
- alergie genetika imunologie mikrobiologie patologie MeSH
- Aspergillus imunologie MeSH
- aspergilóza genetika imunologie mikrobiologie patologie MeSH
- epitelové buňky imunologie mikrobiologie patologie MeSH
- interleukin-17 nedostatek genetika imunologie MeSH
- lidé MeSH
- modely nemocí na zvířatech MeSH
- myši inbrední C57BL MeSH
- myši knockoutované MeSH
- myši MeSH
- náchylnost k nemoci MeSH
- plíce imunologie mikrobiologie patologie MeSH
- protein - isoformy nedostatek genetika imunologie MeSH
- pseudomonádové infekce genetika imunologie mikrobiologie patologie MeSH
- Pseudomonas imunologie MeSH
- receptory interleukinu-17 nedostatek genetika imunologie MeSH
- regulace genové exprese MeSH
- respirační sliznice imunologie mikrobiologie patologie MeSH
- signální transdukce MeSH
- stafylokokové infekce genetika imunologie mikrobiologie patologie MeSH
- Staphylococcus aureus imunologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
γ-Tubulin is the key protein for microtubule nucleation. Duplication of the γ-tubulin gene occurred several times during evolution, and in mammals γ-tubulin genes encode proteins which share ∼97% sequence identity. Previous analysis of Tubg1 and Tubg2 knock-out mice has suggested that γ-tubulins are not functionally equivalent. Tubg1 knock-out mice died at the blastocyst stage, whereas Tubg2 knock-out mice developed normally and were fertile. It was proposed that γ-tubulin 1 represents ubiquitous γ-tubulin, while γ-tubulin 2 may have some specific functions and cannot substitute for γ-tubulin 1 deficiency in blastocysts. The molecular basis of the suggested functional difference between γ-tubulins remains unknown. Here we show that exogenous γ-tubulin 2 is targeted to centrosomes and interacts with γ-tubulin complex proteins 2 and 4. Depletion of γ-tubulin 1 by RNAi in U2OS cells causes impaired microtubule nucleation and metaphase arrest. Wild-type phenotype in γ-tubulin 1-depleted cells is restored by expression of exogenous mouse or human γ-tubulin 2. Further, we show at both mRNA and protein levels using RT-qPCR and 2D-PAGE, respectively, that in contrast to Tubg1, the Tubg2 expression is dramatically reduced in mouse blastocysts. This indicates that γ-tubulin 2 cannot rescue γ-tubulin 1 deficiency in knock-out blastocysts, owing to its very low amount. The combined data suggest that γ-tubulin 2 is able to nucleate microtubules and substitute for γ-tubulin 1. We propose that mammalian γ-tubulins are functionally redundant with respect to the nucleation activity.
- MeSH
- časové faktory MeSH
- down regulace MeSH
- embryonální vývoj genetika MeSH
- implantace embrya MeSH
- intracelulární prostor metabolismus MeSH
- lidé MeSH
- mikrotubuly metabolismus MeSH
- mitóza genetika MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- protein - isoformy nedostatek genetika metabolismus MeSH
- transport proteinů MeSH
- tubulin nedostatek genetika metabolismus MeSH
- vývojová regulace genové exprese MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
