IndraLab

"Given that several DUBs are recruited to their substrates indirectly through binding to the E3 ligase xref , we analyzed whether KLHL2 mediated the interaction between UCK1 and USP28 in vivo ."

"USP28 does not bind directly with UCK1, but binds via KLHL2 [ xref ]."

"Our data strongly suggest that KLHL2 mediates the interaction between UCK1 and USP28."

"IP and IB analyses suggested interactions between KLHL2, UCK1, and USP28."

"USP28 binds to UCK1 via KLHL2."

"Given that several DUBs are recruited to their substrates indirectly through binding to the E3 ligase XREF_BIBR, we analyzed whether KLHL2 mediated the interaction between UCK1 and USP28 in vivo."

"Conversely, silencing KLHL2 significantly decreased the amount of endogenous UCK1 that was bound to HA-USP28 (XREF_SUPPLEMENTARY B)."

"Our data strongly suggest that KLHL2 mediates the interaction between UCK1 and USP28."

"The findings that support this conclusion are the following : (i) the binding of UCK1 to the E3 ligase KLHL2 causes its polyubiquitination at K81 and degradation; (ii) the DUB USP28 binds to UCK1 through interaction with KLHL2 and antagonizes KLHL2 mediated effect on UCK1; (iii) UCK1 deubiquitination by USP28 is dependent on the phosphorylation status of USP28; (iv) USP28 is phosphorylated by 5 '-AZA-activated ATM, resulting in disassociation of KLHL2 from USP28 and UCK1 destabilization; (v) ATM also phosphorylates UCK1 at S145, significantly enhancing the KLHL2 and UCK1 complex formation; (vi) the knockdown of KLHL2 not only significantly inhibits AML cell proliferation, but also sensitizes AML cells to 5 '-AZA-induced apoptosis; and (vii) combinational therapies with 5 '-AZA and ectopic USP28 expression enhances the effects of 5 '-AZA and induces synergistic lethality at cellular level and in the AML mouse model, which is, however, observed in cells deficient in USP28."

"Although KLHL2 and UCK1 formed binary complexes, USP28 did not directly bind to UCK1."