Modulation of LSD1 phosphorylation by CK2/WIP1 regulates RNF168-dependent 53BP1 recruitment in response to DNA damage

Abstract

Proper DNA damage response is essential for the maintenance of genome integrity. The E3 ligase RNF168 deficiency fully prevents both the initial recruitment and retention of 53BP1 at sites of DNA damage. In response to DNA damage, RNF168-dependent recruitment of the lysine-specific demethylase LSD1 to the site of DNA damage promotes local H3K4me2 demethylation and ubiquitination of H2A/H2AX, facilitating 53BP1 recruitment to sites of DNA damage. Alternatively, RNF168-mediated K63-linked ubiquitylation of 53BP1 is required for the initial recruitment of 53BP1 to sites of DNA damage and for its function in repair. We demonstrated here that phosphorylation and dephosphorylation of LSD1 at S131 and S137 was mediated by casein kinase 2 (CK2) and wild-type p53-induced phosphatase 1 (WIP1), respectively. LSD1, RNF168 and 53BP1 interacted with each other directly. CK2-mediated phosphorylation of LSD1 exhibited no impact on its interaction with 53BP1, but promoted its interaction with RNF168 and RNF168-dependent 53BP1 ubiquitination and subsequent recruitment to the DNA damage sites. Furthermore, overexpression of phosphorylation-defective mutants failed to restore LSD1 depletion-induced cellular sensitivity to DNA damage. Taken together, our results suggest that LSD1 phosphorylation modulated by CK2/WIP1 regulates RNF168-dependent 53BP1 recruitment directly in response to DNA damage and cellular sensitivity to DNA damaging agents.

Phosphorylation of LSD1 by PLK1 promotes its chromatin release during mitosis

“…4b). Consistent with the mass spectrometric analysis result, mutation of Ser-126 to Ala fully abolished PLK1-mediated phosphorylation on LSD1, while mutation of the adjacent Ser-131 to Ala, which is a CK2-mediated phosphorylation site [12], did not have a similar effect (Fig. 4a).…”

“…Both immunoblotting and immunoprecipitation assays were performed with desired antibodies according to the protocols described before [12, 21]. …”

“…For in vitro kinase assays [12, 22], bacterially-purified HIS tagged PLK1 and LSD1 were incubated at kinase buffer containing 50 mM HEPES (pH 7.4), 10 mM MgCl 2 , 1 mM DTT, 1 mM Na 3 VO 4 , 10 μM coldATP and 5 μCi [γ- 32 P]-ATP at 30 °C for 30 min.…”

Lysine-Specific Histone Demethylases Contribute to Cellular Differentiation and Carcinogenesis

“…Structurally, KDM1A comprises four domains: an N-terminal disordered segment, a SWIRM (SWI3p, Rsc8p, and Moira) domain, an amine oxidase domain (AOD), and a Tower domain ( Figure 2B). The N-terminal disordered segment contains about 150 amino acids and is subjected to post-translational modifications [13][14][15][16]. This domain is required for KDM1A nuclear localization and protein-protein interactions [17,18].…”

“…In this process, KDM1A interacts with the zinc-finger protein SNAIL1 and demethylates H3K4me1/me2 at epithelial-specific gene promoters, including that of E-cadherin. The loss of H3K4 methylation represses epithelial-specific gene expression, with a consequent transition to the mesenchymal state [13,15,38,39].…”