Mutations in ERK2 binding sites affect nuclear entry

Mustafa N. Yazicioglu, Daryl L. Goad, Aarati Ranganathan, Angelique W. Whitehurst, Elizabeth J. Goldsmith, Melanie H. Cobb

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Abstract

The MAPK ERK2 can enter and exit the nucleus by an energy-independent process that is facilitated by direct interactions with nuclear pore proteins. Several studies also suggest that the localization of ERK2 can be influenced by carrier proteins. Using import reconstitution assays, we examined a group of ERK2 mutants defective in known protein interactions to determine structural properties of ERK2 that contribute to its nuclear entry. ERK2 mutants defective in binding to substrates near the active site or to basic/hydrophobic docking (D) motifs were imported normally. Several ERK2 mutants defective in interactions with FXF motifs displayed slowed rates of nuclear import. The import-impaired mutants also showed reduced binding to a recombinant C-terminal fragment of nucleoporin 153 that is rich in FXF motifs. Despite the deficit revealed in some mutants via reconstitution assays, all but one of the ERK2 mutants accumulated in nuclei of stimulated cells in a manner comparable with the wild type protein; the mutant most defective in import remained in the cytoplasm. These results further support the idea that direct interactions with nucleoporins are involved in ERK2 nuclear entry and that multiple events contribute to the ligand-dependent relocalization of these protein kinases.

Original languageEnglish (US)
Pages (from-to)28759-28767
Number of pages9
JournalJournal of Biological Chemistry
Volume282
Issue number39
DOIs
StatePublished - Sep 28 2007

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Nuclear Pore Complex Proteins
Assays
Binding Sites
Nuclear Pore
Porins
Mutation
Cell Nucleus Active Transport
Nuclear Proteins
Cell Nucleus
Protein Kinases
Structural properties
Catalytic Domain
Carrier Proteins
Cytoplasm
Proteins
Ligands
Substrates

ASJC Scopus subject areas

  • Biochemistry

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Mutations in ERK2 binding sites affect nuclear entry. / Yazicioglu, Mustafa N.; Goad, Daryl L.; Ranganathan, Aarati; Whitehurst, Angelique W.; Goldsmith, Elizabeth J.; Cobb, Melanie H.

In: Journal of Biological Chemistry, Vol. 282, No. 39, 28.09.2007, p. 28759-28767.

Research output: Contribution to journalArticle

Yazicioglu, Mustafa N. ; Goad, Daryl L. ; Ranganathan, Aarati ; Whitehurst, Angelique W. ; Goldsmith, Elizabeth J. ; Cobb, Melanie H. / Mutations in ERK2 binding sites affect nuclear entry. In: Journal of Biological Chemistry. 2007 ; Vol. 282, No. 39. pp. 28759-28767.
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