Equal segregation of chromosomes during mitosis ensures euploidy of daughter cells. Defects in this process may result in an imbalance in the chromosomal composition and cellular transformation. Proteolytic and non-proteolytic ubiquitylation pathways ensure directionality and fidelity of mitotic progression but specific mitotic functions of deubiquitylating enzymes (DUBs) remain less studied. Here we describe the role of the DUB ubiquitin carboxyl-terminal hydrolase isozyme L3 (UCHL3) in the regulation of chromosome bi-orientation and segregation during mitosis. Downregulation or inhibition of UCHL3 leads to chromosome alignment defects during metaphase. Frequent segregation errors during anaphase are also observed upon inactivation of UCHL3. Mechanistically, UCHL3 interacts with and deubiquitylates Aurora B, the catalytic subunit of chromosome passenger complex (CPC), known to be critically involved in the regulation of chromosome alignment and segregation. UCHL3 does not regulate protein levels of Aurora B or the binding of Aurora B to other CPC subunits. Instead, UCHL3 promotes localization of Aurora B to kinetochores, suggesting its role in the error correction mechanism monitoring bi-orientation of chromosomes during metaphase. Thus, UCHL3 contributes to the regulation of faithful genome segregation and maintenance of euploidy in human cells.

• MeSH
• aurora kinasa B fyziologie   MeSH
• HeLa buňky MeSH
• lidé MeSH
• mitóza * MeSH
• segregace chromozomů * MeSH
• thiolesterasa ubikvitinu fyziologie   MeSH
• ubikvitinace MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Reduced nitric oxide (NO)/cGMP signalling is observed in age-related vascular disease. We hypothesize that this disturbed signalling involves effects of genomic instability, a primary causal factor in aging, on vascular smooth muscle cells (VSMCs) and that the underlying mechanism plays a role in human age-related vascular disease. To test our hypothesis, we combined experiments in mice with genomic instability resulting from the defective nucleotide excision repair gene ERCC1 (Ercc1(d/-) mice), human VSMC cultures and population genome-wide association studies (GWAS). Aortic rings of Ercc1(d/-) mice showed 43% reduced responses to the soluble guanylate cyclase (sGC) stimulator sodium nitroprusside (SNP). Inhibition of phosphodiesterase (PDE) 1 and 5 normalized SNP-relaxing effects in Ercc1(d/-) to wild-type (WT) levels. PDE1C levels were increased in lung and aorta. cGMP hydrolysis by PDE in lungs was higher in Ercc1(d/-) mice. No differences in activity or levels of cGMP-dependent protein kinase 1 or sGC were observed in Ercc1(d/-) mice compared with WT. Senescent human VSMC showed elevated PDE1A and PDE1C and PDE5 mRNA levels (11.6-, 9- and 2.3-fold respectively), which associated with markers of cellular senescence. Conversely, PDE1 inhibition lowered expression of these markers. Human genetic studies revealed significant associations of PDE1A single nucleotide polymorphisms with diastolic blood pressure (DBP; β=0.28, P=2.47×10(-5)) and carotid intima-media thickness (cIMT; β=-0.0061, P=2.89×10(-5)). In summary, these results show that genomic instability and cellular senescence in VSMCs increase PDE1 expression. This might play a role in aging-related loss of vasodilator function, VSMC senescence, increased blood pressure and vascular hypertrophy.

• MeSH
• arteriae carotides enzymologie   patologie   MeSH
• celogenomová asociační studie MeSH
• cyklické nukleotidfosfodiesterasy, typ 1 antagonisté a inhibitory   genetika   metabolismus   MeSH
• cyklické nukleotidfosfodiesterasy, typ 5 genetika   metabolismus   MeSH
• DNA vazebné proteiny nedostatek   genetika   MeSH
• endonukleasy nedostatek   genetika   MeSH
• fenotyp MeSH
• genetická predispozice k nemoci MeSH
• guanosinmonofosfát cyklický metabolismus   MeSH
• hydrolýza MeSH
• hyperplazie MeSH
• hypertenze enzymologie   genetika   patofyziologie   MeSH
• inhibitory fosfodiesterasy 5 farmakologie   MeSH
• intimomediální šíře tepenné stěny MeSH
• jednonukleotidový polymorfismus MeSH
• krevní tlak MeSH
• kultivované buňky MeSH
• lidé MeSH
• myocyty hladké svaloviny účinky léků   enzymologie   MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• nemoci arterie carotis enzymologie   genetika   patologie   MeSH
• regulace genové exprese u nádorů MeSH
• stárnutí buněk MeSH
• stárnutí genetika   metabolismus   MeSH
• svaly hladké cévní účinky léků   enzymologie   MeSH
• systémy druhého messengeru MeSH
• techniky in vitro MeSH
• vazodilatace * účinky léků   MeSH
• vazodilatancia farmakologie   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

We previously identified genomic instability as a causative factor for vascular aging. In the present study, we determined which vascular aging outcomes are due to local endothelial DNA damage, which was accomplished by genetic removal of ERCC1 (excision repair cross-complementation group 1) DNA repair in mice (EC-knockout (EC-KO) mice). EC-KO showed a progressive decrease in microvascular dilation of the skin, increased microvascular leakage in the kidney, decreased lung perfusion, and increased aortic stiffness compared with wild-type (WT). EC-KO showed expression of DNA damage and potential senescence marker p21 exclusively in the endothelium, as demonstrated in aorta. Also the kidney showed p21-positive cells. Vasodilator responses measured in organ baths were decreased in aorta, iliac and coronary artery EC-KO compared with WT, of which coronary artery was the earliest to be affected. Nitric oxide-mediated endothelium-dependent vasodilation was abolished in aorta and coronary artery, whereas endothelium-derived hyperpolarization and responses to exogenous nitric oxide (NO) were intact. EC-KO showed increased superoxide production compared with WT, as measured in lung tissue, rich in endothelial cells (ECs). Arterial systolic blood pressure (BP) was increased at 3 months, but normal at 5 months, at which age cardiac output (CO) was decreased. Since no further signs of cardiac dysfunction were detected, this decrease might be an adaptation to prevent an increase in BP. In summary, a selective DNA repair defect in the endothelium produces features of age-related endothelial dysfunction, largely attributed to loss of endothelium-derived NO. Increased superoxide generation might contribute to the observed changes affecting end organ perfusion, as demonstrated in kidney and lung.

• MeSH
• cévní endotel metabolismus   patologie   patofyziologie   MeSH
• DNA vazebné proteiny nedostatek   genetika   MeSH
• endonukleasy nedostatek   genetika   MeSH
• endoteliální buňky metabolismus   patologie   MeSH
• inhibitor p21 cyklin-dependentní kinasy genetika   metabolismus   MeSH
• kapilární permeabilita MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• oprava DNA * MeSH
• oxid dusnatý metabolismus   MeSH
• poškození DNA * MeSH
• stárnutí buněk genetika   MeSH
• stárnutí genetika   metabolismus   patologie   MeSH
• superoxidy metabolismus   MeSH
• synthasa oxidu dusnatého, typ III metabolismus   MeSH
• tuhost cévní stěny MeSH
• vazodilatace MeSH
• věkové faktory MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

• MeSH
• adalimumab MeSH
• biologická terapie MeSH
• biosimilární léčivé přípravky MeSH
• humanizované monoklonální protilátky MeSH
• idiopatické střevní záněty farmakoterapie   MeSH
• klinické lékařství MeSH
• TNF-alfa antagonisté a inhibitory   MeSH
• Publikační typ
• přehledy MeSH

• Konspekt
• Farmacie. Farmakologie
• NLK Obory
• farmacie a farmakologie
• gastroenterologie