Karyopherin-α2 Protein Interacts with Chk2 and Contributes to Its Nuclear Import

Laura Zannini, Daniele Lecis, Sofia Lisanti, Roberta Benetti, Giacomo Buscemi, Claudio Schneider, Domenico Delia

Research output: Contribution to journalArticlepeer-review

Abstract

Chk2 is a nuclear protein kinase involved in the DNA damage-induced ataxia telangiectasia mutated-dependent checkpoint arrest at multiple cell cycle phases. Searching for Chk2-binding proteins by a yeast two-hybrid system, we identified a strong interaction with karyopherin-α2 (KPNA-2), a gene product involved in active nuclear import of proteins bearing a nuclear localization signal (NLS). This finding was confirmed by glutathione S-transferase pull-down and co-immunoprecipitation assays. Of the three predicted Chk2 NLSs, located at amino acids 179-182 (NLS-1), 240-256 (NLS-2), and 515-522 (NLS-3), only the latter mediated the interaction with KPNA-2 in the yeast two-hybrid system, and in particular with its C terminus. Unlike mutations in NLS-1 or NLS-2, which left the nuclear localization of Chk2 unaffected, mutations in NLS-3 caused a cytoplasmic relocalization, indicating that the NLS-3 motif acts indeed as NLS for Chk2 in vivo. Finally, co-transfection experiments with green fluorescent protein (GFP)-Chk2 and wild type or mutant KPNA-2 confirmed the role of KPNA-2 in nuclear import of Chk2.

Original languageEnglish
Pages (from-to)42346-42351
Number of pages6
JournalJournal of Biological Chemistry
Volume278
Issue number43
DOIs
Publication statusPublished - Oct 24 2003

ASJC Scopus subject areas

  • Biochemistry

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