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Nat Commun. 2014 Mar 6;5:3394. doi: 10.1038/ncomms4394.

Tousled-like kinases phosphorylate Asf1 to promote histone supply during DNA replication.

Author information

1
1] Biotech Research and Innovation Centre (BRIC) and Centre for Epigenetics, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen DK-2200, Denmark [2].
2
1] Department of Biochemistry & Molecular Biology and Centre for Epigenetics, University of Southern Denmark, Campusvej 55, Odense DK-5230, Denmark [2] Department of Proteomics, The Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Blegdamsvej 3B, Copenhagen DK-2200, Denmark.
3
Biotech Research and Innovation Centre (BRIC) and Centre for Epigenetics, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen DK-2200, Denmark.
4
The Finsen Laboratory, Rigshospitalet, Copenhagen Biocenter, Ole Maaløes Vej 5, Copenhagen DK-2200, Denmark.
5
Department of Proteomics, The Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Blegdamsvej 3B, Copenhagen DK-2200, Denmark.
6
Department of Biochemistry & Molecular Biology and Centre for Epigenetics, University of Southern Denmark, Campusvej 55, Odense DK-5230, Denmark.

Abstract

During DNA replication, nucleosomes are rapidly assembled on newly synthesized DNA to restore chromatin organization. Asf1, a key histone H3-H4 chaperone required for this process, is phosphorylated by Tousled-like kinases (TLKs). Here, we identify TLK phosphorylation sites by mass spectrometry and dissect how phosphorylation has an impact on human Asf1 function. The divergent C-terminal tail of Asf1a is phosphorylated at several sites, and this is required for timely progression through S phase. Consistent with this, biochemical analysis of wild-type and phospho-mimetic Asf1a shows that phosphorylation enhances binding to histones and the downstream chaperones CAF-1 and HIRA. Moreover, we find that TLK phosphorylation of Asf1a is induced in cells experiencing deficiency of new histones and that TLK interaction with Asf1a involves its histone-binding pocket. We thus propose that TLK signalling promotes histone supply in S phase by targeting histone-free Asf1 and stimulating its ability to shuttle histones to sites of chromatin assembly.

PMID:
24598821
PMCID:
PMC3977046
DOI:
10.1038/ncomms4394
[Indexed for MEDLINE]
Free PMC Article
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