Synthetic Lethal Mutations with spmex67 of Schizosaccharomyces pombe in the Mediation of mRNA Export

  • Yoon, Jin-Ho (Department of Biology, Sungshin Women′s University)
  • Published : 2003.06.01

Abstract

Mex67p/Tap are evolutionally conserved mRNA export factors. To identify mutations in genes that are functionally linked to mex67 with respect to mRNA export, we used a synthetic lethal genetic screen in Schizosaccharomyces pombe. Three synthetic lethal mutants were isolated and mutations in these mutants defined separate complementation groups. These mutants exhibited the accumulation of poly A$\^$+/ RNA in the nucleus, with a decrease in the cytoplasm under synthetically lethal conditions, suggesting that the mutations cause an mRNA nuclear export defect. In addition, the S. pombe genes that were found to be involved in mRNA export did not suppress the synthetic lethality of these mutants. These results indicate that the isolated mutants contain mutations in new genes, which are involved in mRNA export from the nucleus.

Keywords

References

  1. Experiments with fission yeast Alfa,C.;P.Fantes;J.Hyams;M.Mcleod;E.Warbrick
  2. Genes Dev. v.6 Isolation and characterization of RAT1: an essential gene of Saccharomyces cerevisiae required for the efficient nucleocytoplasmic trafficking of mRNA Amberg,D.C.;A.L.Goldstein;C.N.Cole https://doi.org/10.1101/gad.6.7.1173
  3. Gene v.123 TATA box mutations in the Schizosaccharomyces pombe nmt1 promoter affect transcription efficiency but not the transcription start point or thiamine repressibility Basi,G.;E.Schmid;K.Maundrell https://doi.org/10.1016/0378-1119(93)90552-E
  4. J. Biol. Chem. v.270 A mutation in the Schizosaccharomyces pombe rae1 gene causes defects in poly(A)+ RNA export and in the cytoskeleton Brown,J.A.;A.Bharathi;A.Ghosh;W.Whalen;E.Fitzgerald;R.Dhar https://doi.org/10.1074/jbc.270.13.7411
  5. Curr. Opin. Cell. Biol. v.13 Nucleocytoplasmic transport enters the atomic age Conti,E.;E.Izaurralde https://doi.org/10.1016/S0955-0674(00)00213-1
  6. Trends Genet. v.11 Genetic approaches to nuclear pore structure and function Doye,V.;E.C.Hurt https://doi.org/10.1016/S0168-9525(00)89057-5
  7. Nat. Rev. Mol. Cell. Biol. v.3 Messenger-RNA-binding proteins and the messages they carry Dreyfuss,G.;V.N.Kim;N.Kataoka https://doi.org/10.1038/nrm760
  8. Annu. Rev. Genet. v.31 Yeast genetics to dissect the nuclear pore complex and nucleocytoplasmic trafficking Fabre,E.;E.Hurt https://doi.org/10.1146/annurev.genet.31.1.277
  9. Nucleic Acids Res. v.21 Comparison of Schizosaccharomyces pombe expression systems Forsburg,S.L. https://doi.org/10.1093/nar/21.12.2955
  10. J. Biol. Chem. v.277 Elf1p, a member of the ABC class of ATPases, functions as a mRNA export factor in Schizosacchromyces pombe Kozak,L.;G.Gopal;J.H.Yoon;Z.E.Sauna;S.V.Ambudkar;A.G.Thakurta;R.Dhar https://doi.org/10.1074/jbc.M205415200
  11. Dev. Cell. v.2 Protein and RNA export from the nucleus Lei,E.P.;P.A.Silver https://doi.org/10.1016/S1534-5807(02)00134-X
  12. Nature v.416 An extensive network of coupling among gene expression machines Maniatis,T.;R.Reed https://doi.org/10.1038/416499a
  13. Gene v.123 Thiamine-repressible expression vectors pREP and pRIP for fission yeast Maundrell,K. https://doi.org/10.1016/0378-1119(93)90551-D
  14. Methods Enzymol. v.194 Molecular genetic analysis of fission yeast Schizosaccharomyces pombe Moreno,S.;A.Klar;P.Nurse https://doi.org/10.1016/0076-6879(91)94059-L
  15. Cell v.99 Transport of proteins and RNAs in and out of the nucleus Nakielny,S.;G.Dreyfuss https://doi.org/10.1016/S0092-8674(00)81666-9
  16. J. Cell. Biol. v.145 RAE1 is a shuttling mRNA export factor that binds to a GLEBS-like NUP98 motif at the nuclear pore complex through multiple domains Pritchard,C.E.;M.Fornerod;L.H.Kasper;J.M. van Deursen https://doi.org/10.1083/jcb.145.2.237
  17. Cell v.108 A conserved mRNA export machinery coupled to pre-mRNA splicing Reed,R.;E.Hurt https://doi.org/10.1016/S0092-8674(02)00627-X
  18. Nat. Cell. Biol. v.3 A new view of mRNA export: separating the wheat from the chaff Reed,R.;K.Magni https://doi.org/10.1038/ncb0901-e201
  19. J. Biol. Chem. v.276 The nuclear pore complex as a transport machine Rout,M.P.;J.D.Aitchison https://doi.org/10.1074/jbc.R100015200
  20. EMBO J. v.16 Mex67p, a novel factor for nuclear mRNA export, binds to both poly(A)+ RNA and nuclear pores Segref,A.;K.Sharma;V.Doye;A.Hellwig;J.Huber;R.Luhrmann;E.Hurt https://doi.org/10.1093/emboj/16.11.3256
  21. Curr. Opin. Cell Biol. v.11 The nuclear pore complex: from molecular architecture to functional dynamics Stoffler,D.;B.Fahrenkrog;U.Aebi https://doi.org/10.1016/S0955-0674(99)80055-6
  22. J. Cell. Biol. v.150 Binding of the Mex67p/Mtr2p heterodimer to FXFG, GLFG, and FG repeat nucleoporins is essential for nuclear mRNA export Strasser,K.;J.Bassler;E.Hurt https://doi.org/10.1083/jcb.150.4.695
  23. EMBO J. v.19 Yra1p, a conserved nuclear RNA-binding protein, interacts directly with Mex67p and is required for mRNA export Strasser,K.;E.Hurt https://doi.org/10.1093/emboj/19.3.410
  24. Nature v.413 Splicing factor Sub2p is required for nuclear mRNA export through its interaction with Yra1p Strasser,K.;E.Hurt https://doi.org/10.1038/35098113
  25. J. Biol. Chem. v.276 The GLFG regions of Nup116p and Nup100p serve as binding sites for both Kap95p and Mex67p at the nuclear pore complex Strawn,L.A.;T.Shen;S.R.Wente https://doi.org/10.1074/jbc.M008311200
  26. Genes Dev. v.12 Nuclear RNA export Stutz,F.;M.Rosbash https://doi.org/10.1101/gad.12.21.3303
  27. Mol. Biol. Cell v.13 Crp79p, like Mex67p, is an auxiliary mRNA export factor in Schizosaccharomyces pombe Thakurta,A.G.;W.A.Whalen;J.H.Yoon;A.Bharathi;L.Kozak;C.Whiteford;D.C.Love;J.A.Hanover;R.Dhar https://doi.org/10.1091/mbc.E01-11-0133
  28. Curr. Opin. Cell Biol. v.13 Nuclear pores and nuclear assembly Vasu,S.K.;D.J.Forbes https://doi.org/10.1016/S0955-0674(00)00221-0
  29. Curr. Opin. Cell Biol. v.14 Nucleocytoplasmic transport: cargo trafficking across the border Weis,K. https://doi.org/10.1016/S0955-0674(02)00337-X
  30. Cell v.112 Regulating Access to the Genome. Nucleocytoplasmic Transport throughout the Cell Cycle Weis,K. https://doi.org/10.1016/S0092-8674(03)00082-5
  31. Genetics v.152 Regulation of mRNA export by nutritional status in fission yeast Whalen,W.A.;J.H.Yoon;R.Shen;R.Dhar
  32. Mol. Cell Biol. v.20 Mex67p of Schizosaccharomyces pombe interacts with Rae1p in mediating mRNA export Yoon,J.H.;D.C.Love;A.Guhathakurta;J.A.Hanover;R.Dhar https://doi.org/10.1128/MCB.20.23.8767-8782.2000
  33. Mol. Cell Biol. v.17 Npp106p, a Schizosaccharomyces pombe nucleoporin similar to Saccharomyces cerevisiae Nic96p, functionally interacts with Rae1p in mRNA export Yoon,J.H.;W.A.Whalen;A.Bharathi;R.Shen;R.Dhar
  34. FEBS Lett. v.498 Nuclear export of mRNA Zenklusen,D.;F.Stutz https://doi.org/10.1016/S0014-5793(01)02482-6