Abstract
The regulation of chromatin structure is critical for a wide range of essential cellular processes. The Tousled-like kinases, TLK1 and TLK2, regulate ASF1, a histone H3/H4 chaperone, and likely other substrates, and their activity has been implicated in transcription, DNA replication, DNA repair, RNA interference, cell cycle progression, viral latency, chromosome segregation and mitosis. However, little is known about the functions of TLK activity in vivo or the relative functions of the highly similar TLK1 and TLK2 in any cell type. To begin to address this, we have generated Tlk1- and Tlk2-deficient mice. We found that while TLK1 was dispensable for murine viability, TLK2 loss led to late embryonic lethality because of placental failure. TLK2 was required for normal trophoblast differentiation and the phosphorylation of ASF1 was reduced in placentas lacking TLK2. Conditional bypass of the placental phenotype allowed the generation of apparently healthy Tlk2-deficient mice, while only the depletion of both TLK1 and TLK2 led to extensive genomic instability, indicating that both activities contribute to genome maintenance. Our data identifies a specific role for TLK2 in placental function during mammalian development and suggests that TLK1 and TLK2 have largely redundant roles in genome maintenance.
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Acknowledgements
We are grateful to the Stracker lab, I del Barco, A Igea, J Roig and C Gekas for helpful input; A Nebreda for critical reading of the manuscript; R Medema for sharing unpublished results; S Aivio for animal maintenance; D Reverter and P Gallego Alonso for purified LC8 protein; J Gloeckner for SF-TAP vectors; the IRB Histopathology facility and N Prats, M Aguilera, B Domínguez, E Llonch and N Vasconcelos for assistance; the IRB Advanced Digital Microscopy Core Facility for use of the Lightsheet Macroscope; L Bardia, M Marsal and S Tosi for sample preparation and image analysis; and E Sabido for technical advice and mass spectrometry measurements. Analysis was performed in the CRG/UPF Proteomics Unit, part of the 'Plataforma de Recursos Biomoleculares y Bioinformàticos' (ProteoRed-Instituto de Salud Carlos III, PT13/0001). THS was a Ramon y Cajal scholar and supported by the Ministerio de Economía y Competitividad (MINECO) (BFU2012-39521, BFU2015-68354, Ayudas para incentivar la incorporación estable de doctores (IED) 2015) and institutional funding from MINECO through the Centres of Excellence Severo Ochoa award and from the CERCA Programme of the Catalan Government. AG is an EMBO Young Investigator, and supported by the Danish National Research Foundation to the Center for Epigenetics (DNRF82), the European Commission ITN FP7 'aDDRess', a European Research Council Starting Grant (ERC2011StG, no. 281 765), the Danish Cancer Society, the Danish Medical Research Council, the Novo Nordisk Foundation and the Lundbeck Foundation. SSB and HG were supported by PhD fellowships from Fundacio La Caixa, PAK was supported by an Advanced Postdoc Mobility fellowship from the Swiss National Science Foundation and the Kurt und Senta Herrmann Stiftung and KR was supported by an FPI fellowship (MINECO).
Author contributions
SSB, PAK and HG designed, performed and analyzed most of the experiments; HG and AP carried out IP-MS experiments; PAK, EC and BR carried out BioID experiments; SAY and AdB assessed placental pathology; KR performed experiments; LP and PAK maintained mouse cohorts; JC performed lightsheet imaging; AG, TLR and SF generated and provided critical reagents and experimental advice; and THS designed and analyzed experiments and wrote the manuscript.
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Segura-Bayona, S., Knobel, P., González-Burón, H. et al. Differential requirements for Tousled-like kinases 1 and 2 in mammalian development. Cell Death Differ 24, 1872–1885 (2017). https://doi.org/10.1038/cdd.2017.108
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DOI: https://doi.org/10.1038/cdd.2017.108
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