Interaction between Ran and Mog1 is Required for Efficient Nuclear Protein Import

Rosanna Baker, Michelle T. Harreman, John F. Eccleston, Anita H. Corbett, Murray Stewart

Research output: Contribution to journalArticle

Abstract

Mog1 is a nuclear protein that interacts with Ran, the Ras family GTPase that confers directionality to nuclear import and export pathways. Deletion of MOG1 in Saccharomyces cerevisiae (Δmog1) causes temperature-sensitive growth and defects in nuclear protein import. Mog1 has previously been shown to stimulate GTP release from Ran and we demonstrate here that addition of Mog1 to either Ran-GTP or Ran-GDP results in nucleotide release and formation of a stable complex between Mog1 and nucleotide-free Ran. Moreover, MOG1 shows synthetic lethality with PRP20, the Ran guanine nucleotide exchange factor (RanGEF) that also binds nucleotide-free Ran. To probe the functional role of the Mog1-Ran interaction, we engineered mutants of yeast Mog1 and Ran that specifically disrupt their interaction both in vitro and in vivo. These mutants indicate that the interaction interface involves conserved Mog1p residues Asp62 and Glu65, and residue Lys136 in yeast Ran. Mutations at these residues decrease the ability of Mog1 to bind and release nucleotide from Ran. Furthermore, the E65K-Mog1 and K136E-Ran mutations in yeast cause temperature sensitivity and mislocalization of a nuclear import reporter protein, similar to the phenotype observed for the Δmog1 strain. Our results indicate that a primary function of Mog1 requires binding to Ran and that the Mog1-Ran interaction is necessary for efficient nuclear protein import in vivo.

Original languageEnglish (US)
Pages (from-to)41255-41262
Number of pages8
JournalJournal of Biological Chemistry
Volume276
Issue number44
DOIs
StatePublished - Nov 2 2001
Externally publishedYes

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Cell Nucleus Active Transport
Nuclear Proteins
Yeast
Nucleotides
Guanosine Triphosphate
Yeasts
Guanine Nucleotide Exchange Factors
GTP Phosphohydrolases
ras Proteins
Mutation
Temperature
Saccharomyces cerevisiae
Defects
Phenotype
Growth
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Interaction between Ran and Mog1 is Required for Efficient Nuclear Protein Import. / Baker, Rosanna; Harreman, Michelle T.; Eccleston, John F.; Corbett, Anita H.; Stewart, Murray.

In: Journal of Biological Chemistry, Vol. 276, No. 44, 02.11.2001, p. 41255-41262.

Research output: Contribution to journalArticle

Baker, Rosanna ; Harreman, Michelle T. ; Eccleston, John F. ; Corbett, Anita H. ; Stewart, Murray. / Interaction between Ran and Mog1 is Required for Efficient Nuclear Protein Import. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 44. pp. 41255-41262.
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