title: Tousled-like kinases phosphorylate Asf1 to promote histone supply during DNA replication.
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.
authors.affiliation: 1] Biotech Research and Innovation Centre (BRIC) and Centre for Epigenetics, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen DK-2200, Denmark [2].
authors.forename: Ilnaz M
authors.initials: IM
authors.lastname: Klimovskaia
authors.affiliation: 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.
authors.forename: Clifford
authors.initials: C
authors.lastname: Young
authors.affiliation: Biotech Research and Innovation Centre (BRIC) and Centre for Epigenetics, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen DK-2200, Denmark.
authors.forename: Caroline B
authors.initials: CB
authors.lastname: Strømme
authors.affiliation: 1] Biotech Research and Innovation Centre (BRIC) and Centre for Epigenetics, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen DK-2200, Denmark [2].
authors.forename: Patrice
authors.initials: P
authors.lastname: Menard
authors.affiliation: Biotech Research and Innovation Centre (BRIC) and Centre for Epigenetics, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen DK-2200, Denmark.
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Jasencakova
authors.affiliation: 1] Biotech Research and Innovation Centre (BRIC) and Centre for Epigenetics, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen DK-2200, Denmark [2].
authors.forename: Jakob
authors.initials: J
authors.lastname: Mejlvang
authors.affiliation: Biotech Research and Innovation Centre (BRIC) and Centre for Epigenetics, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen DK-2200, Denmark.
authors.forename: Katrine
authors.initials: K
authors.lastname: Ask
authors.affiliation: The Finsen Laboratory, Rigshospitalet, Copenhagen Biocenter, Ole Maaløes Vej 5, Copenhagen DK-2200, Denmark.
authors.forename: Michael
authors.initials: M
authors.lastname: Ploug
authors.affiliation: Department of Proteomics, The Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Blegdamsvej 3B, Copenhagen DK-2200, Denmark.
authors.forename: Michael L
authors.initials: ML
authors.lastname: Nielsen
authors.affiliation: Department of Biochemistry & Molecular Biology and Centre for Epigenetics, University of Southern Denmark, Campusvej 55, Odense DK-5230, Denmark.
authors.forename: Ole N
authors.initials: ON
authors.lastname: Jensen
authors.affiliation: Biotech Research and Innovation Centre (BRIC) and Centre for Epigenetics, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen DK-2200, Denmark.
authors.forename: Anja
authors.initials: A
authors.lastname: Groth
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D015153:Blotting, Western; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D004261:DNA Replication; D006367:HeLa Cells; D006657:Histones / Q000378:metabolism*; D006801:Humans; D013058:Mass Spectrometry; D018613:Microscopy, Confocal; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D010766:Phosphorylation; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D016196:S Phase / Q000235:genetics
chemical_list: C491641:ASF1A protein, human; C491640:ASF1B protein, human; D018797:Cell Cycle Proteins; D002843:Chromatin; D006657:Histones; D018832:Molecular Chaperones; D017346:Protein Serine-Threonine Kinases; C425323:TLK1 protein, human