Nuclear translocation promotes proteasomal degradation of human Rad17 protein through the N-terminal destruction boxes

Abstract

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Rad17 Translocates to Nucleolus upon UV Irradiation through Nucleolar Localization Signal in the Central Basic Domain

“…The K/R359–363A mutant of this construct was deficient in the accumulation in the nucleolus and equally distributed in the nucleolus and the nucleoplasm. This mutation also increased the cytoplasmic localization (42%, Figure 2 A,B and Figure S1A,B ) relative to WT, as was shown recently [ 14 ]. EGFP alone was equally distributed in the nucleolus and the nucleoplasm, and no specific localization in the nucleoplasm or the cytoplasm was observed ( Figure 2 A,B and Figure S1A ).…”

“…In our recent study, we found that EGFP fused with Rad17 E295–D380 peptide showed exclusive nuclear localization [ 14 ]. In this study, we further characterized the central basic domain of Rad17 spanning N339–D380 ( Figure 1 A).…”

“…In our previous work, we showed that Rad17 K359–K363 encoded a part of the nuclear localization signal and that K359A/R360A/R361A/K362A/K363A (K/R359–363A or 5KR) mutation abolished the nuclear localization of Rad17 [ 14 ]. Here, we examined the effect of K/R359–363A mutation on the nucleolar localization.…”

“…We have recently reported that the central basic domain contains a nuclear localization signal and that the nuclear translocation of Rad17 promotes its proteasomal degradation. We have also identified tandem destruction boxes of Rad17 on its N-terminus as a set of canonical and noncanonical sequences [ 14 ]. The proteasomal degradation of Rad17 is mediated by an anaphase-promoting complex associated with Cdh1 [ 8 ], and the N-terminal destruction boxes of Rad17 interact with Cdh1 in vitro [ 14 ].…”

“…We have also identified tandem destruction boxes of Rad17 on its N-terminus as a set of canonical and noncanonical sequences [ 14 ]. The proteasomal degradation of Rad17 is mediated by an anaphase-promoting complex associated with Cdh1 [ 8 ], and the N-terminal destruction boxes of Rad17 interact with Cdh1 in vitro [ 14 ].…”