IndraLab

Statements

PLK2 phosphorylates NPM1. 11 / 11
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"We show that Plk2 interacts with and phosphorylates NPM and B23 on S4 in both mitosis and S-phase while Plk1 seems to be involved only in its mitotic phosphorylation."
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"Furthermore, we demonstrate that Plk2 dependent phosphorylation of NPM and B23 on S4 triggers both centriole reduplication and normal centrosome reproduction."
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"To further verify that NPM and B23 is phosphorylated by Plk2 on S4 we used a phospho specific antibody against NPM and B23 phosphorylated on S4 (pS4 NPM and B23) in a reaction where His tagged NPM and B23 WT or NPM and B23 S4A was incubated with Plk2 WT or KD in an in vitro kinase assay."
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"But NPM1 is also phosphorylated at S4 by Plk1 during mitosis and by Plk-2 near the G1/S transition."
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"More recently, it has been shown that NPM is also downstream of other signaling pathways, as phosphorylation of NPM by Plk2 is critical to centrosome duplication [ xref ]."
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"While [gamma 32 P] was efficiently incorporated in both NPM and B23 WT and the T199A mutant, the NPM and B23 S4A mutant was not phosphorylated by Plk2 (XREF_FIG) confirming that S4 is the site that is phosphorylated by Plk2."
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"Furthermore, to address whether Plk2 could phosphorylate NPM and B23, we carried out in vitro kinase reactions."
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"XREF_FIG shows that the NPM and B23 S4A-mutant can not be phosphorylated by active Plk2."
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"Together, these results show that NPM and B23 is phosphorylated on S4 by both Plk2 and Plk1 in vitro."
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"To directly confirm that NPM and B23 is phosphorylated by Plk2 in S-phase HeLa cells were transfected with either Flag tagged Plk2 WT or Plk2 KD and then treated with hydroxyurea (HU) to arrest cells in early S-phase."
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"The results showed that Plk2 phosphorylates NPM and B23 in vitro."
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