title: Phosphorylation of KRAB-associated protein 1 (KAP1) at Tyr-449, Tyr-458, and Tyr-517 by nuclear tyrosine kinases inhibits the association of KAP1 and heterochromatin protein 1α (HP1α) with heterochromatin.
abstract: Protein tyrosine phosphorylation regulates a wide range of cellular processes at the plasma membrane. Recently, we showed that nuclear tyrosine phosphorylation by Src family kinases (SFKs) induces chromatin structural changes. In this study, we identify KRAB-associated protein 1 (KAP1/TIF1β/TRIM28), a component of heterochromatin, as a nuclear tyrosine-phosphorylated protein. Tyrosine phosphorylation of KAP1 is induced by several tyrosine kinases, such as Src, Lyn, Abl, and Brk. Among SFKs, Src strongly induces tyrosine phosphorylation of KAP1. Nucleus-targeted Lyn potentiates tyrosine phosphorylation of KAP1 compared with intact Lyn, but neither intact Fyn nor nucleus-targeted Fyn phosphorylates KAP1. Substitution of the three tyrosine residues Tyr-449/Tyr-458/Tyr-517, located close to the HP1 binding-motif, into phenylalanine ablates tyrosine phosphorylation of KAP1. Immunostaining and chromatin fractionation show that Src and Lyn decrease the association of KAP1 with heterochromatin in a kinase activity-dependent manner. KAP1 knockdown impairs the association of HP1α with heterochromatin, because HP1α associates with KAP1 in heterochromatin. Intriguingly, tyrosine phosphorylation of KAP1 decreases the association of HP1α with heterochromatin, which is inhibited by replacement of endogenous KAP1 with its phenylalanine mutant (KAP1-Y449F/Y458F/Y517F, KAP1-3YF). In DNA damage, KAP1-3YF repressed transcription of p21. These results suggest that nucleus-localized tyrosine kinases, including SFKs, phosphorylate KAP1 at Tyr-449/Tyr-458/Tyr-517 and inhibit the association of KAP1 and HP1α with heterochromatin.
authors: Department of Molecular Cell Biology, Graduate School of Pharmaceutical Sciences, Chiba University, Inohana 1-8-1, Chuo-ku, Chiba 260-8675, Japan.
authors: Sho
authors: S
authors: Kubota
authors: Yasunori
authors: Y
authors: Fukumoto
authors: Kazumasa
authors: K
authors: Aoyama
authors: Kenichi
authors: K
authors: Ishibashi
authors: Ryuzaburo
authors: R
authors: Yuki
authors: Takao
authors: T
authors: Morinaga
authors: Takuya
authors: T
authors: Honda
authors: Noritaka
authors: N
authors: Yamaguchi
authors: Takahisa
authors: T
authors: Kuga
authors: Takeshi
authors: T
authors: Tomonaga
authors: Naoto
authors: N
authors: Yamaguchi
mesh_terms: D000818:Animals; D019556:COS Cells; D002467:Cell Nucleus / Q000201:enzymology; D002522:Chlorocebus aethiops; D000090266:Chromobox Protein Homolog 5; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism*; D006367:HeLa Cells; D006570:Heterochromatin / Q000378:metabolism*; D006801:Humans; D010766:Phosphorylation; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D012097:Repressor Proteins / Q000378:metabolism*; D000076183:Tripartite Motif-Containing Protein 28; D014443:Tyrosine / Q000378:metabolism; D019061:src-Family Kinases / Q000378:metabolism*
chemical_list: C000715594:CBX5 protein, human; D002868:Chromosomal Proteins, Non-Histone; D006570:Heterochromatin; D012097:Repressor Proteins; D000090266:Chromobox Protein Homolog 5; D014443:Tyrosine; C101116:TRIM28 protein, human; D000076183:Tripartite Motif-Containing Protein 28; D011505:Protein-Tyrosine Kinases; D019061:src-Family Kinases
keywords: HP1; Heterochromatin; KAP1; Nucleus; Protein Phosphorylation; Protein-Protein Interactions; Tyrosine Protein Kinase (Tyrosine Kinase)
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