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. 1999 May 4;18(9):2593–2609. doi: 10.1093/emboj/18.9.2593

The Mex67p-mediated nuclear mRNA export pathway is conserved from yeast to human.

J Katahira 1, K Strässer 1, A Podtelejnikov 1, M Mann 1, J U Jung 1, E Hurt 1
PMCID: PMC1171339  PMID: 10228171

Abstract

Human TAP is an orthologue of the yeast mRNA export factor Mex67p. In mammalian cells, TAP has a preferential intranuclear localization, but can also be detected at the nuclear pores and shuttles between the nucleus and the cytoplasm. TAP directly associates with mRNA in vivo, as it can be UV-crosslinked to poly(A)+ RNA in HeLa cells. Both the FG-repeat domain of nucleoporin CAN/Nup214 and a novel human 15 kDa protein (p15) with homology to NTF2 (a nuclear transport factor which associates with RanGDP), directly bind to TAP. When green fluorescent protein (GFP)-tagged TAP and p15 are expressed in yeast, they localize to the nuclear pores. Strikingly, co-expression of human TAP and p15 restores growth of the otherwise lethal mex67::HIS3/mtr2::HIS3 double knockout strain. Thus, the human TAP-p15 complex can functionally replace the Mex67p-Mtr2p complex in yeast and thus performs a conserved role in nuclear mRNA export.

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Selected References

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  1. Aris J. P., Blobel G. cDNA cloning and sequencing of human fibrillarin, a conserved nucleolar protein recognized by autoimmune antisera. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):931–935. doi: 10.1073/pnas.88.3.931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arts G. J., Fornerod M., Mattaj I. W. Identification of a nuclear export receptor for tRNA. Curr Biol. 1998 Mar 12;8(6):305–314. doi: 10.1016/s0960-9822(98)70130-7. [DOI] [PubMed] [Google Scholar]
  3. Bastos R., Lin A., Enarson M., Burke B. Targeting and function in mRNA export of nuclear pore complex protein Nup153. J Cell Biol. 1996 Sep;134(5):1141–1156. doi: 10.1083/jcb.134.5.1141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Belgareh N., Snay-Hodge C., Pasteau F., Dagher S., Cole C. N., Doye V. Functional characterization of a Nup159p-containing nuclear pore subcomplex. Mol Biol Cell. 1998 Dec;9(12):3475–3492. doi: 10.1091/mbc.9.12.3475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Boer J. M., van Deursen J. M., Croes H. J., Fransen J. A., Grosveld G. C. The nucleoporin CAN/Nup214 binds to both the cytoplasmic and the nucleoplasmic sides of the nuclear pore complex in overexpressing cells. Exp Cell Res. 1997 Apr 10;232(1):182–185. doi: 10.1006/excr.1997.3502. [DOI] [PubMed] [Google Scholar]
  6. Bogerd H. P., Fridell R. A., Madore S., Cullen B. R. Identification of a novel cellular cofactor for the Rev/Rex class of retroviral regulatory proteins. Cell. 1995 Aug 11;82(3):485–494. doi: 10.1016/0092-8674(95)90437-9. [DOI] [PubMed] [Google Scholar]
  7. Borer R. A., Lehner C. F., Eppenberger H. M., Nigg E. A. Major nucleolar proteins shuttle between nucleus and cytoplasm. Cell. 1989 Feb 10;56(3):379–390. doi: 10.1016/0092-8674(89)90241-9. [DOI] [PubMed] [Google Scholar]
  8. Bullock T. L., Clarkson W. D., Kent H. M., Stewart M. The 1.6 angstroms resolution crystal structure of nuclear transport factor 2 (NTF2). J Mol Biol. 1996 Jul 19;260(3):422–431. doi: 10.1006/jmbi.1996.0411. [DOI] [PubMed] [Google Scholar]
  9. Carmo-Fonseca M., Kern H., Hurt E. C. Human nucleoporin p62 and the essential yeast nuclear pore protein NSP1 show sequence homology and a similar domain organization. Eur J Cell Biol. 1991 Jun;55(1):17–30. [PubMed] [Google Scholar]
  10. Clarkson W. D., Kent H. M., Stewart M. Separate binding sites on nuclear transport factor 2 (NTF2) for GDP-Ran and the phenylalanine-rich repeat regions of nucleoporins p62 and Nsp1p. J Mol Biol. 1996 Nov 8;263(4):517–524. doi: 10.1006/jmbi.1996.0594. [DOI] [PubMed] [Google Scholar]
  11. Conti E., Uy M., Leighton L., Blobel G., Kuriyan J. Crystallographic analysis of the recognition of a nuclear localization signal by the nuclear import factor karyopherin alpha. Cell. 1998 Jul 24;94(2):193–204. doi: 10.1016/s0092-8674(00)81419-1. [DOI] [PubMed] [Google Scholar]
  12. Dahlberg J. E., Lund E. Functions of the GTPase Ran in RNA export from the nucleus. Curr Opin Cell Biol. 1998 Jun;10(3):400–408. doi: 10.1016/s0955-0674(98)80017-3. [DOI] [PubMed] [Google Scholar]
  13. Del Priore V., Heath C., Snay C., MacMillan A., Gorsch L., Dagher S., Cole C. A structure/function analysis of Rat7p/Nup159p, an essential nucleoporin of Saccharomyces cerevisiae. J Cell Sci. 1997 Dec;110(Pt 23):2987–2999. doi: 10.1242/jcs.110.23.2987. [DOI] [PubMed] [Google Scholar]
  14. Doye V., Wepf R., Hurt E. C. A novel nuclear pore protein Nup133p with distinct roles in poly(A)+ RNA transport and nuclear pore distribution. EMBO J. 1994 Dec 15;13(24):6062–6075. doi: 10.1002/j.1460-2075.1994.tb06953.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Dreyfuss G., Matunis M. J., Piñol-Roma S., Burd C. G. hnRNP proteins and the biogenesis of mRNA. Annu Rev Biochem. 1993;62:289–321. doi: 10.1146/annurev.bi.62.070193.001445. [DOI] [PubMed] [Google Scholar]
  16. Fabre E., Hurt E. Yeast genetics to dissect the nuclear pore complex and nucleocytoplasmic trafficking. Annu Rev Genet. 1997;31:277–313. doi: 10.1146/annurev.genet.31.1.277. [DOI] [PubMed] [Google Scholar]
  17. Fornerod M., Ohno M., Yoshida M., Mattaj I. W. CRM1 is an export receptor for leucine-rich nuclear export signals. Cell. 1997 Sep 19;90(6):1051–1060. doi: 10.1016/s0092-8674(00)80371-2. [DOI] [PubMed] [Google Scholar]
  18. Fornerod M., van Deursen J., van Baal S., Reynolds A., Davis D., Murti K. G., Fransen J., Grosveld G. The human homologue of yeast CRM1 is in a dynamic subcomplex with CAN/Nup214 and a novel nuclear pore component Nup88. EMBO J. 1997 Feb 17;16(4):807–816. doi: 10.1093/emboj/16.4.807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Fukuda M., Asano S., Nakamura T., Adachi M., Yoshida M., Yanagida M., Nishida E. CRM1 is responsible for intracellular transport mediated by the nuclear export signal. Nature. 1997 Nov 20;390(6657):308–311. doi: 10.1038/36894. [DOI] [PubMed] [Google Scholar]
  20. Gabler S., Schütt H., Groitl P., Wolf H., Shenk T., Dobner T. E1B 55-kilodalton-associated protein: a cellular protein with RNA-binding activity implicated in nucleocytoplasmic transport of adenovirus and cellular mRNAs. J Virol. 1998 Oct;72(10):7960–7971. doi: 10.1128/jvi.72.10.7960-7971.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Grüter P., Tabernero C., von Kobbe C., Schmitt C., Saavedra C., Bachi A., Wilm M., Felber B. K., Izaurralde E. TAP, the human homolog of Mex67p, mediates CTE-dependent RNA export from the nucleus. Mol Cell. 1998 Apr;1(5):649–659. doi: 10.1016/s1097-2765(00)80065-9. [DOI] [PubMed] [Google Scholar]
  22. Hellmuth K., Lau D. M., Bischoff F. R., Künzler M., Hurt E., Simos G. Yeast Los1p has properties of an exportin-like nucleocytoplasmic transport factor for tRNA. Mol Cell Biol. 1998 Nov;18(11):6374–6386. doi: 10.1128/mcb.18.11.6374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Huber J., Cronshagen U., Kadokura M., Marshallsay C., Wada T., Sekine M., Lührmann R. Snurportin1, an m3G-cap-specific nuclear import receptor with a novel domain structure. EMBO J. 1998 Jul 15;17(14):4114–4126. doi: 10.1093/emboj/17.14.4114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Hurwitz M. E., Strambio-de-Castillia C., Blobel G. Two yeast nuclear pore complex proteins involved in mRNA export form a cytoplasmically oriented subcomplex. Proc Natl Acad Sci U S A. 1998 Sep 15;95(19):11241–11245. doi: 10.1073/pnas.95.19.11241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Jensen O. N., Podtelejnikov A., Mann M. Delayed extraction improves specificity in database searches by matrix-assisted laser desorption/ionization peptide maps. Rapid Commun Mass Spectrom. 1996;10(11):1371–1378. doi: 10.1002/(SICI)1097-0231(199608)10:11<1371::AID-RCM682>3.0.CO;2-5. [DOI] [PubMed] [Google Scholar]
  26. Katahira J., Inoue N., Horiguchi Y., Matsuda M., Sugimoto N. Molecular cloning and functional characterization of the receptor for Clostridium perfringens enterotoxin. J Cell Biol. 1997 Mar 24;136(6):1239–1247. doi: 10.1083/jcb.136.6.1239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kraemer D. M., Strambio-de-Castillia C., Blobel G., Rout M. P. The essential yeast nucleoporin NUP159 is located on the cytoplasmic side of the nuclear pore complex and serves in karyopherin-mediated binding of transport substrate. J Biol Chem. 1995 Aug 11;270(32):19017–19021. doi: 10.1074/jbc.270.32.19017. [DOI] [PubMed] [Google Scholar]
  28. Kraemer D., Wozniak R. W., Blobel G., Radu A. The human CAN protein, a putative oncogene product associated with myeloid leukemogenesis, is a nuclear pore complex protein that faces the cytoplasm. Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1519–1523. doi: 10.1073/pnas.91.4.1519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kutay U., Bischoff F. R., Kostka S., Kraft R., Görlich D. Export of importin alpha from the nucleus is mediated by a specific nuclear transport factor. Cell. 1997 Sep 19;90(6):1061–1071. doi: 10.1016/s0092-8674(00)80372-4. [DOI] [PubMed] [Google Scholar]
  30. Kutay U., Lipowsky G., Izaurralde E., Bischoff F. R., Schwarzmaier P., Hartmann E., Görlich D. Identification of a tRNA-specific nuclear export receptor. Mol Cell. 1998 Feb;1(3):359–369. doi: 10.1016/s1097-2765(00)80036-2. [DOI] [PubMed] [Google Scholar]
  31. Künzler M., Hurt E. C. Cse1p functions as the nuclear export receptor for importin alpha in yeast. FEBS Lett. 1998 Aug 21;433(3):185–190. doi: 10.1016/s0014-5793(98)00892-8. [DOI] [PubMed] [Google Scholar]
  32. Mann M., Wilm M. Error-tolerant identification of peptides in sequence databases by peptide sequence tags. Anal Chem. 1994 Dec 15;66(24):4390–4399. doi: 10.1021/ac00096a002. [DOI] [PubMed] [Google Scholar]
  33. Mattaj I. W., Englmeier L. Nucleocytoplasmic transport: the soluble phase. Annu Rev Biochem. 1998;67:265–306. doi: 10.1146/annurev.biochem.67.1.265. [DOI] [PubMed] [Google Scholar]
  34. Matunis E. L., Matunis M. J., Dreyfuss G. Characterization of the major hnRNP proteins from Drosophila melanogaster. J Cell Biol. 1992 Jan;116(2):257–269. doi: 10.1083/jcb.116.2.257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Matunis M. J., Matunis E. L., Dreyfuss G. PUB1: a major yeast poly(A)+ RNA-binding protein. Mol Cell Biol. 1993 Oct;13(10):6114–6123. doi: 10.1128/mcb.13.10.6114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Michael W. M., Choi M., Dreyfuss G. A nuclear export signal in hnRNP A1: a signal-mediated, temperature-dependent nuclear protein export pathway. Cell. 1995 Nov 3;83(3):415–422. doi: 10.1016/0092-8674(95)90119-1. [DOI] [PubMed] [Google Scholar]
  37. Nakielny S., Dreyfuss G. Import and export of the nuclear protein import receptor transportin by a mechanism independent of GTP hydrolysis. Curr Biol. 1998 Jan 15;8(2):89–95. doi: 10.1016/s0960-9822(98)70039-9. [DOI] [PubMed] [Google Scholar]
  38. Nakielny S., Dreyfuss G. The hnRNP C proteins contain a nuclear retention sequence that can override nuclear export signals. J Cell Biol. 1996 Sep;134(6):1365–1373. doi: 10.1083/jcb.134.6.1365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Neville M., Stutz F., Lee L., Davis L. I., Rosbash M. The importin-beta family member Crm1p bridges the interaction between Rev and the nuclear pore complex during nuclear export. Curr Biol. 1997 Oct 1;7(10):767–775. doi: 10.1016/s0960-9822(06)00335-6. [DOI] [PubMed] [Google Scholar]
  40. Ohno M., Fornerod M., Mattaj I. W. Nucleocytoplasmic transport: the last 200 nanometers. Cell. 1998 Feb 6;92(3):327–336. doi: 10.1016/s0092-8674(00)80926-5. [DOI] [PubMed] [Google Scholar]
  41. Ossareh-Nazari B., Bachelerie F., Dargemont C. Evidence for a role of CRM1 in signal-mediated nuclear protein export. Science. 1997 Oct 3;278(5335):141–144. doi: 10.1126/science.278.5335.141. [DOI] [PubMed] [Google Scholar]
  42. Percipalle P., Clarkson W. D., Kent H. M., Rhodes D., Stewart M. Molecular interactions between the importin alpha/beta heterodimer and proteins involved in vertebrate nuclear protein import. J Mol Biol. 1997 Mar 7;266(4):722–732. doi: 10.1006/jmbi.1996.0801. [DOI] [PubMed] [Google Scholar]
  43. Piñol-Roma S., Adam S. A., Choi Y. D., Dreyfuss G. Ultraviolet-induced cross-linking of RNA to proteins in vivo. Methods Enzymol. 1989;180:410–418. doi: 10.1016/0076-6879(89)80114-4. [DOI] [PubMed] [Google Scholar]
  44. Piñol-Roma S., Dreyfuss G. Shuttling of pre-mRNA binding proteins between nucleus and cytoplasm. Nature. 1992 Feb 20;355(6362):730–732. doi: 10.1038/355730a0. [DOI] [PubMed] [Google Scholar]
  45. Piñol-Roma S., Swanson M. S., Gall J. G., Dreyfuss G. A novel heterogeneous nuclear RNP protein with a unique distribution on nascent transcripts. J Cell Biol. 1989 Dec;109(6 Pt 1):2575–2587. doi: 10.1083/jcb.109.6.2575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Powers M. A., Forbes D. J., Dahlberg J. E., Lund E. The vertebrate GLFG nucleoporin, Nup98, is an essential component of multiple RNA export pathways. J Cell Biol. 1997 Jan 27;136(2):241–250. doi: 10.1083/jcb.136.2.241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Reichelt R., Holzenburg A., Buhle E. L., Jr, Jarnik M., Engel A., Aebi U. Correlation between structure and mass distribution of the nuclear pore complex and of distinct pore complex components. J Cell Biol. 1990 Apr;110(4):883–894. doi: 10.1083/jcb.110.4.883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Rexach M., Blobel G. Protein import into nuclei: association and dissociation reactions involving transport substrate, transport factors, and nucleoporins. Cell. 1995 Dec 1;83(5):683–692. doi: 10.1016/0092-8674(95)90181-7. [DOI] [PubMed] [Google Scholar]
  49. Santos-Rosa H., Moreno H., Simos G., Segref A., Fahrenkrog B., Panté N., Hurt E. Nuclear mRNA export requires complex formation between Mex67p and Mtr2p at the nuclear pores. Mol Cell Biol. 1998 Nov;18(11):6826–6838. doi: 10.1128/mcb.18.11.6826. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Schneider C., Newman R. A., Sutherland D. R., Asser U., Greaves M. F. A one-step purification of membrane proteins using a high efficiency immunomatrix. J Biol Chem. 1982 Sep 25;257(18):10766–10769. [PubMed] [Google Scholar]
  51. Segref A., Sharma K., Doye V., Hellwig A., Huber J., Lührmann R., Hurt E. Mex67p, a novel factor for nuclear mRNA export, binds to both poly(A)+ RNA and nuclear pores. EMBO J. 1997 Jun 2;16(11):3256–3271. doi: 10.1093/emboj/16.11.3256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Shevchenko A., Chernushevich I., Ens W., Standing K. G., Thomson B., Wilm M., Mann M. Rapid 'de novo' peptide sequencing by a combination of nanoelectrospray, isotopic labeling and a quadrupole/time-of-flight mass spectrometer. Rapid Commun Mass Spectrom. 1997;11(9):1015–1024. doi: 10.1002/(SICI)1097-0231(19970615)11:9<1015::AID-RCM958>3.0.CO;2-H. [DOI] [PubMed] [Google Scholar]
  53. Shevchenko A., Jensen O. N., Podtelejnikov A. V., Sagliocco F., Wilm M., Vorm O., Mortensen P., Shevchenko A., Boucherie H., Mann M. Linking genome and proteome by mass spectrometry: large-scale identification of yeast proteins from two dimensional gels. Proc Natl Acad Sci U S A. 1996 Dec 10;93(25):14440–14445. doi: 10.1073/pnas.93.25.14440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Shevchenko A., Wilm M., Vorm O., Mann M. Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal Chem. 1996 Mar 1;68(5):850–858. doi: 10.1021/ac950914h. [DOI] [PubMed] [Google Scholar]
  55. Siniossoglou S., Wimmer C., Rieger M., Doye V., Tekotte H., Weise C., Emig S., Segref A., Hurt E. C. A novel complex of nucleoporins, which includes Sec13p and a Sec13p homolog, is essential for normal nuclear pores. Cell. 1996 Jan 26;84(2):265–275. doi: 10.1016/s0092-8674(00)80981-2. [DOI] [PubMed] [Google Scholar]
  56. Solsbacher J., Maurer P., Bischoff F. R., Schlenstedt G. Cse1p is involved in export of yeast importin alpha from the nucleus. Mol Cell Biol. 1998 Nov;18(11):6805–6815. doi: 10.1128/mcb.18.11.6805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Stade K., Ford C. S., Guthrie C., Weis K. Exportin 1 (Crm1p) is an essential nuclear export factor. Cell. 1997 Sep 19;90(6):1041–1050. doi: 10.1016/s0092-8674(00)80370-0. [DOI] [PubMed] [Google Scholar]
  58. Stewart M., Kent H. M., McCoy A. J. Structural basis for molecular recognition between nuclear transport factor 2 (NTF2) and the GDP-bound form of the Ras-family GTPase Ran. J Mol Biol. 1998 Apr 3;277(3):635–646. doi: 10.1006/jmbi.1997.1602. [DOI] [PubMed] [Google Scholar]
  59. Stutz F., Neville M., Rosbash M. Identification of a novel nuclear pore-associated protein as a functional target of the HIV-1 Rev protein in yeast. Cell. 1995 Aug 11;82(3):495–506. doi: 10.1016/0092-8674(95)90438-7. [DOI] [PubMed] [Google Scholar]
  60. Stutz F., Rosbash M. Nuclear RNA export. Genes Dev. 1998 Nov 1;12(21):3303–3319. doi: 10.1101/gad.12.21.3303. [DOI] [PubMed] [Google Scholar]
  61. Ullman K. S., Powers M. A., Forbes D. J. Nuclear export receptors: from importin to exportin. Cell. 1997 Sep 19;90(6):967–970. doi: 10.1016/s0092-8674(00)80361-x. [DOI] [PubMed] [Google Scholar]
  62. Wente S. R., Rout M. P., Blobel G. A new family of yeast nuclear pore complex proteins. J Cell Biol. 1992 Nov;119(4):705–723. doi: 10.1083/jcb.119.4.705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Wilm M., Mann M. Analytical properties of the nanoelectrospray ion source. Anal Chem. 1996 Jan 1;68(1):1–8. doi: 10.1021/ac9509519. [DOI] [PubMed] [Google Scholar]
  64. Wilm M., Shevchenko A., Houthaeve T., Breit S., Schweigerer L., Fotsis T., Mann M. Femtomole sequencing of proteins from polyacrylamide gels by nano-electrospray mass spectrometry. Nature. 1996 Feb 1;379(6564):466–469. doi: 10.1038/379466a0. [DOI] [PubMed] [Google Scholar]
  65. Yang Q., Rout M. P., Akey C. W. Three-dimensional architecture of the isolated yeast nuclear pore complex: functional and evolutionary implications. Mol Cell. 1998 Jan;1(2):223–234. doi: 10.1016/s1097-2765(00)80023-4. [DOI] [PubMed] [Google Scholar]
  66. Yoon D. W., Lee H., Seol W., DeMaria M., Rosenzweig M., Jung J. U. Tap: a novel cellular protein that interacts with tip of herpesvirus saimiri and induces lymphocyte aggregation. Immunity. 1997 May;6(5):571–582. doi: 10.1016/s1074-7613(00)80345-3. [DOI] [PubMed] [Google Scholar]
  67. van Deursen J., Boer J., Kasper L., Grosveld G. G2 arrest and impaired nucleocytoplasmic transport in mouse embryos lacking the proto-oncogene CAN/Nup214. EMBO J. 1996 Oct 15;15(20):5574–5583. [PMC free article] [PubMed] [Google Scholar]

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