Maintenance of genome stability is essential for every living cell as genetic information is repeatedly challenged during DNA replication in each cell division event. Errors, defects, delays, and mistakes that arise during mitosis or meiosis lead to an activation of DNA repair processes and in case of their failure, programmed cell death, i.e. apoptosis, could be initiated. Fam208a is a protein whose importance in heterochromatin maintenance has been described recently. In this work, we describe the crucial role of Fam208a in sustaining the genome stability during the cellular division. The targeted depletion of Fam208a in mice using CRISPR/Cas9 leads to embryonic lethality before E12.5. We also used the siRNA approach to downregulate Fam208a in zygotes to avoid the influence of maternal RNA in the early stages of development. This early downregulation increased arresting of the embryonal development at the two-cell stage and occurrence of multipolar spindles formation. To investigate this further, we used the yeast two-hybrid (Y2H) system and identified new putative interaction partners Gpsm2, Amn1, Eml1, Svil, and Itgb3bp. Their co-expression with Fam208a was assessed by qRT-PCR profiling and in situ hybridisation [1] in multiple murine tissues. Based on these results we proposed that Fam208a functions within the HUSH complex by interaction with Mphosph8 as these proteins are not only able to physically interact but also co-localise. We are bringing new evidence that Fam208a is multi-interacting protein affecting genome stability on the level of cell division at the earliest stages of development and also by interaction with methylation complex in adult tissues. In addition to its epigenetic functions, Fam208a appears to have an additional role in zygotic division, possibly via interaction with newly identified putative partners Gpsm2, Amn1, Eml1, Svil, and Itgb3bp.

• MeSH
• aparát dělícího vřeténka metabolismus   MeSH
• buněčné dělení genetika   fyziologie   MeSH
• CRISPR-Cas systémy MeSH
• embryonální vývoj genetika   fyziologie   MeSH
• fosfoproteiny metabolismus   MeSH
• HEK293 buňky MeSH
• jaderné proteiny fyziologie   MeSH
• letální geny MeSH
• lidé MeSH
• malá interferující RNA genetika   farmakologie   MeSH
• multiproteinové komplexy MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• myši MeSH
• nestabilita genomu MeSH
• RNA interference MeSH
• vývojová regulace genové exprese * MeSH
• zvířata MeSH
• zygota metabolismus   MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• imunita MeSH
• imunitní systém fyziologie   patofyziologie   MeSH
• imunoterapie MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• alergologie a imunologie
• biologie
• fyziologie
• NLK Publikační typ
• kolektivní monografie

The protein kinases ataxia-telangiectasia mutated (ATM) and ATM-Rad3 related (ATR) are activated in response to DNA damage, genotoxic stress and virus infections. Here we show that during infection with wild-type adenovirus, ATR and its cofactors RPA32, ATRIP and TopBP1 accumulate at viral replication centres, but there is minimal ATR activation. We show that the Mre11/Rad50/Nbs1 (MRN) complex is recruited to viral centres only during infection with adenoviruses lacking the early region E4 and ATR signaling is activated. This suggests a novel requirement for the MRN complex in ATR activation during virus infection, which is independent of Mre11 nuclease activity and recruitment of RPA/ATR/ATRIP/TopBP1. Unlike other damage scenarios, we found that ATM and ATR signaling are not dependent on each other during infection. We identify a region of the viral E4orf3 protein responsible for immobilization of the MRN complex and show that this prevents ATR signaling during adenovirus infection. We propose that immobilization of the MRN damage sensor by E4orf3 protein prevents recognition of viral genomes and blocks detrimental aspects of checkpoint signaling during virus infection.

• MeSH
• Adenoviridae MeSH
• adenovirové infekce * metabolismus   MeSH
• adenovirové proteiny E4 chemie   metabolismus   MeSH
• ATM protein MeSH
• buněčné linie MeSH
• DNA vazebné proteiny * metabolismus   MeSH
• enzymy opravy DNA * metabolismus   MeSH
• fosforylace MeSH
• jaderné proteiny * metabolismus   MeSH
• lidé MeSH
• molekulární sekvence - údaje MeSH
• multiproteinové komplexy * metabolismus   MeSH
• nádorové supresorové proteiny metabolismus   MeSH
• protein-serin-threoninkinasy * metabolismus   MeSH
• proteiny buněčného cyklu * metabolismus   MeSH
• replikace viru MeSH
• sekvence aminokyselin MeSH
• signální transdukce * MeSH
• transport proteinů MeSH
• Check Tag
• lidé MeSH

• MeSH
• biologie buňky MeSH
• molekulární biologie MeSH
• Publikační typ
• monografie MeSH

• Konspekt
• Biochemie. Molekulární biologie. Biofyzika
• NLK Obory
• biologie
• cytologie, klinická cytologie