The nuclear pore complex and the DEAD box protein Rat8p/Dbp5p have nonessential features which appear to facilitate mRNA export following heat shock.
Nuclear pore complexes (NPCs) play an essential role in RNA export. Nucleoporins required for mRNA export in Saccharomyces cerevisiae are found in the Nup84p and Nup82p subcomplexes of the NPC. The Nup82p subcomplex contains Nup82p, Rat7p/Nup159p, Nsp1p, Gle1p/Rss1p, and Rip1p/Nup42p and is found only on the cytoplasmic face of NPCs. Both Rat7p and Gle1p contain binding sites for Rat8p/Dbp5p, an essential DEAD box protein and putative RNA helicase. Rip1p interacts directly with Gle1p and is the only protein known to be essential for mRNA export after heat shock but not under normal growth conditions. We report that in cells lacking Rip1p, both Gle1p and Rat8p dissociate from NPCs following heat shock at 42 degrees C. Rat8p but not Gle1p was retained at NPCs if rip1Delta cells were first shifted to 37 degrees C and then to 42 degrees C, and this was correlated with preserving mRNA export in heat-shocked rip1Delta cells. Export following ethanol shock was less dependent on the presence of Rip1p. Exposure to 10% ethanol led to dissociation of Rat8p from NPCs in both wild-type and rip1Delta cells. Following this treatment, Rat8p was primarily nuclear in wild-type cells but primarily cytoplasmic in rip1Delta cells. We also determined that efficient export of heat shock mRNA after heat shock depends upon a novel 6-amino-acid element within Rat8p. This motif is not required under normal growth conditions or following ethanol shock. These studies suggest that the molecular mechanism responsible for the defect in export of heat shock mRNAs in heat-shocked rip1Delta cells is dissociation of Rat8p from NPCs. These studies also suggest that both nuclear pores and Rat8p have features not required for mRNA export in growing cells but which enhance the ability of mRNAs to be exported following heat shock.
Rollenhagen
Christiane
C
Department of Biochemistry, Dartmouth Medical School, 7200 Vail Building, Hanover, NH 03755, USA.
Hodge
Christine A
CA
Cole
Charles N
CN
D000891:Anti-Infective Agents, Local / Q000494:pharmacology; D002352:Carrier Proteins / Q000378:metabolism; D053487:DEAD-box RNA Helicases; D000431:Ethanol / Q000494:pharmacology; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics; D006358:Hot Temperature; D022022:Nuclear Pore / Q000378:metabolism*; D028861:Nuclear Pore Complex Proteins; D029543:Nucleocytoplasmic Transport Proteins / Q000378:metabolism*; D020365:RNA Helicases / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D020539:Sequence Analysis, Protein
D000891:Anti-Infective Agents, Local; D002352:Carrier Proteins; C101408:GLE1 protein, S cerevisiae; D018840:HSP70 Heat-Shock Proteins; D028861:Nuclear Pore Complex Proteins; D029543:Nucleocytoplasmic Transport Proteins; D012333:RNA, Messenger; D012261:Ribonucleoproteins; C456269:SSA4 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; C026452:messenger ribonucleoprotein; D000431:Ethanol; C492382:DBP5 protein, S cerevisiae; D053487:DEAD-box RNA Helicases; D020365:RNA Helicases