Rat8p/Dbp5p is a shuttling transport factor that interacts with Rat7p/Nup159p and Gle1p and suppresses the mRNA export defect of xpo1-1 cells

• Published in: The EMBO journal Vol. 18; no. 20; pp. 5778 - 5788
• Main Authors: Hodge, C.A, Colot, H.V, Stafford, P, Cole, C.N
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 15.10.1999
• Subjects: Adenosine Triphosphatases - metabolism; binding proteins; Biological Transport, Active; Carrier Proteins - genetics; Carrier Proteins - metabolism; cell membranes; Cell Nucleus - metabolism; cytochemistry; cytoplasm; Cytoplasm - metabolism; DEAD-box proteins; DEAD-box RNA Helicases; Exportin 1 Protein; Fungal Proteins - metabolism; interactions; Karyopherins; Membrane Proteins - metabolism; messenger RNA; mRNA export; Mutation; Nuclear Pore Complex Proteins; Nuclear Proteins - metabolism; nuclei; nucleocytoplasmic shuttling; Nucleocytoplasmic Transport Proteins; nucleoporins; RAT8; Receptors, Cytoplasmic and Nuclear; RNA Helicases; RNA, Fungal - metabolism; RNA, Messenger - metabolism; RNA-Binding Proteins - metabolism; Saccharomyces cerevisiae; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - growth & development; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins; transfer; YMR255w-GFD1
• ISSN: 0261-4189; 1460-2075; 1460-2075
• DOI: 10.1093/emboj/18.20.5778

In a screen for temperature‐sensitive mutants of Saccharomyces cerevisiae defective for mRNA export, we previously identified the essential DEAD‐box protein Dbp5p/Rat8p and the nucleoporin Rat7p/Nup159p. Both are essential for mRNA export. Here we report that Dbp5p and Rat7p interact through their Nterminal domains. Deletion of this portion of Rat7p (Rat7pΔN) results in strong defects in mRNA export and eliminates association of Dbp5p with nuclear pores. Overexpression of Dbp5p completely suppressed the growth and mRNA export defects of rat7ΔN cells and resulted in weaker suppression in cells carrying rat7‐1 or the rss1‐37 allele of GLE1. Dbp5p interacts with Gle1p independently of the N‐terminus of Dbp5p. Dbp5p shuttles between nucleus and cytoplasm in an Xpo1p‐dependent manner. It accumulates in nuclei of xpo1‐1 cells and in cells with mutations affecting Mex67p (mex67‐5), Gsp1p (Ran) or Ran effectors. Overexpression of Dbp5p prevents nuclear accumulation of mRNA in xpo1‐1 cells, but does not restore growth, suggesting that the RNA export defect of xpo1‐1 cells may be indirect. In a screen for high‐copy suppressors of the rat8‐2 allele of DBP5, we identified YMR255w, now called GFD1. Gfd1p is not essential, interacts with Gle1p and Rip1p/Nup42p, and is found in the cytoplasm and at the nuclear rim.