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

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.

[1]  A. Shevchenko,et al.  Rapid 'de novo' peptide sequencing by a combination of nanoelectrospray, isotopic labeling and a quadrupole/time-of-flight mass spectrometer. , 1997, Rapid communications in mass spectrometry : RCM.

[2]  W. Seol,et al.  Tap: a novel cellular protein that interacts with tip of herpesvirus saimiri and induces lymphocyte aggregation. , 1997, Immunity.

[3]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[4]  M. Carmo-Fonseca,et al.  Human nucleoporin p62 and the essential yeast nuclear pore protein NSP1 show sequence homology and a similar domain organization. , 1991, European journal of cell biology.

[5]  M. Fornerod,et al.  The human homologue of yeast CRM1 is in a dynamic subcomplex with CAN/Nup214 and a novel nuclear pore component Nup88 , 1997, The EMBO journal.

[6]  Mitsuo Sekine,et al.  Snurportin1, an m3G‐cap‐specific nuclear import receptor with a novel domain structure , 1998, The EMBO journal.

[7]  G. Blobel,et al.  Two yeast nuclear pore complex proteins involved in mRNA export form a cytoplasmically oriented subcomplex. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[8]  M. Powers,et al.  The Vertebrate GLFG Nucleoporin, Nup98, Is an Essential Component of Multiple RNA Export Pathways , 1997, The Journal of cell biology.

[9]  C. Dargemont,et al.  Evidence for a role of CRM1 in signal-mediated nuclear protein export. , 1997, Science.

[10]  F. Bischoff,et al.  Export of Importin α from the Nucleus Is Mediated by a Specific Nuclear Transport Factor , 1997, Cell.

[11]  F. Bischoff,et al.  Cse1p Is Involved in Export of Yeast Importin α from the Nucleus , 1998, Molecular and Cellular Biology.

[12]  T. Shenk,et al.  E1B 55-Kilodalton-Associated Protein: a Cellular Protein with RNA-Binding Activity Implicated in Nucleocytoplasmic Transport of Adenovirus and Cellular mRNAs , 1998, Journal of Virology.

[13]  A. McCoy,et al.  Structural basis for molecular recognition between nuclear transport factor 2 (NTF2) and the GDP-bound form of the Ras-family GTPase Ran. , 1998, Journal of molecular biology.

[14]  Minoru Yoshida,et al.  CRM1 is responsible for intracellular transport mediated by the nuclear export signal , 1997, Nature.

[15]  G. Blobel,et al.  A new family of yeast nuclear pore complex proteins , 1992, The Journal of cell biology.

[16]  B. Cullen,et al.  Identification of a novel cellular cofactor for the Rev/Rex class of retroviral regulatory proteins , 1995, Cell.

[17]  R. Lührmann,et al.  Mex67p, a novel factor for nuclear mRNA export, binds to both poly(A)+ RNA and nuclear pores , 1997, The EMBO journal.

[18]  M. Rieger,et al.  A Novel Complex of Nucleoporins, Which Includes Sec13p and a Sec13p Homolog, Is Essential for Normal Nuclear Pores , 1996, Cell.

[19]  Helen M. Kent,et al.  The 1.6 Å Resolution Crystal Structure of Nuclear Transport Factor 2 (NTF2) , 1996 .

[20]  Minoru Yoshida,et al.  CRM1 Is an Export Receptor for Leucine-Rich Nuclear Export Signals , 1997, Cell.

[21]  C. Lehner,et al.  Major nucleolar proteins shuttle between nucleus and cytoplasm , 1989, Cell.

[22]  M. Fornerod,et al.  Identification of a nuclear export receptor for tRNA , 1998, Current Biology.

[23]  G. Dreyfuss,et al.  PUB1: a major yeast poly(A)+ RNA-binding protein. , 1993, Molecular and cellular biology.

[24]  A. Bruno,et al.  Hologram-based refractive index detector for capillary electrophoresis: separation of metal ions , 1994 .

[25]  M. Fornerod,et al.  Nucleocytoplasmic Transport: The Last 200 Nanometers , 1998, Cell.

[26]  E. Lund,et al.  Functions of the GTPase Ran in RNA export from the nucleus. , 1998, Current opinion in cell biology.

[27]  A. Podtelejnikov,et al.  Delayed extraction improves specificity in database searches by matrix-assisted laser desorption/ionization peptide maps. , 1996, Rapid communications in mass spectrometry : RCM.

[28]  J. Fransen,et al.  The nucleoporin CAN/Nup214 binds to both the cytoplasmic and the nucleoplasmic sides of the nuclear pore complex in overexpressing cells. , 1997, Experimental cell research.

[29]  C. Cole,et al.  A structure/function analysis of Rat7p/Nup159p, an essential nucleoporin of Saccharomyces cerevisiae. , 1997, Journal of cell science.

[30]  G. Dreyfuss,et al.  A nuclear export signal in hnRNP A1: A signal-mediated, temperature-dependent nuclear protein export pathway , 1995, Cell.

[31]  E. Hurt,et al.  Yeast genetics to dissect the nuclear pore complex and nucleocytoplasmic trafficking. , 1997, Annual review of genetics.

[32]  G. Dreyfuss,et al.  A novel heterogeneous nuclear RNP protein with a unique distribution on nascent transcripts , 1989, The Journal of cell biology.

[33]  M. Künzler,et al.  Yeast Los1p Has Properties of an Exportin-Like Nucleocytoplasmic Transport Factor for tRNA , 1998, Molecular and Cellular Biology.

[34]  G. Lipowsky,et al.  NTF2 mediates nuclear import of Ran , 1998, The EMBO journal.

[35]  M. Wilm,et al.  Analytical properties of the nanoelectrospray ion source. , 1996, Analytical chemistry.

[36]  C. Cole,et al.  Functional characterization of a Nup159p-containing nuclear pore subcomplex. , 1998, Molecular biology of the cell.

[37]  Helena Santos-Rosa,et al.  Nuclear mRNA Export Requires Complex Formation between Mex67p and Mtr2p at the Nuclear Pores , 1998, Molecular and Cellular Biology.

[38]  R. Wepf,et al.  A novel nuclear pore protein Nup133p with distinct roles in poly(A)+ RNA transport and nuclear pore distribution. , 1994, The EMBO journal.

[39]  M. Künzler,et al.  Cselp functions as the nuclear export receptor for importin α in yeast , 1998 .

[40]  D. Rhodes,et al.  Molecular interactions between the importin α/β heterodimer and proteins involved in vertebrate nuclear protein import , 1997 .

[41]  J. Katahira,et al.  Molecular Cloning and Functional Characterization of the Receptor for Clostridium perfringens Enterotoxin , 1997, The Journal of cell biology.

[42]  D. Görlich Transport into and out of the cell nucleus , 1998, The EMBO journal.

[43]  M. Wilm,et al.  TAP, the human homolog of Mex67p, mediates CTE-dependent RNA export from the nucleus. , 1998, Molecular cell.

[44]  A. Shevchenko,et al.  Femtomole sequencing of proteins from polyacrylamide gels by nano-electrospray mass spectrometry , 1996, Nature.

[45]  G. Dreyfuss,et al.  Ultraviolet-induced cross-linking of RNA to proteins in vivo. , 1989, Methods in enzymology.

[46]  M. Rosbash,et al.  Nuclear RNA export. , 1998, Genes & development.

[47]  M. Wilm,et al.  Error-tolerant identification of peptides in sequence databases by peptide sequence tags. , 1994, Analytical chemistry.

[48]  K. Umesono,et al.  Intracellular Localization and Transcriptional Activation by the Human Glucocorticoid Receptor-Green Fluorescent Protein (GFP) Fusion Proteins , 1998 .

[49]  G. Blobel,et al.  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 (*) , 1995, The Journal of Biological Chemistry.

[50]  M. Rosbash,et al.  The importin-beta family member Crm1p bridges the interaction between Rev and the nuclear pore complex during nuclear export , 1997, Current Biology.

[51]  G. Dreyfuss,et al.  The hnRNP C proteins contain a nuclear retention sequence that can override nuclear export signals , 1996, The Journal of cell biology.

[52]  B. Burke,et al.  Targeting and function in mRNA export of nuclear pore complex protein Nup153 , 1996, The Journal of cell biology.

[53]  C. Akey,et al.  Three-dimensional architecture of the isolated yeast nuclear pore complex: functional and evolutionary implications. , 1998, Molecular cell.

[54]  Utz Fischer,et al.  The HIV-1 Rev Activation Domain is a nuclear export signal that accesses an export pathway used by specific cellular RNAs , 1995, Cell.

[55]  G. Dreyfuss,et al.  Import and export of the nuclear protein import receptor transportin by a mechanism independent of GTP hydrolysis , 1998, Current Biology.

[56]  Karsten Weis,et al.  Exportin 1 (Crm1p) Is an Essential Nuclear Export Factor , 1997, Cell.

[57]  G. Dreyfuss,et al.  Shuttling of pre-mRNA binding proteins between nucleus and cytoplasm , 1992, Nature.

[58]  G. Blobel,et al.  Protein import into nuclei: association and dissociation reactions involving transport substrate, transport factors, and nucleoporins , 1995, Cell.

[59]  G. Blobel,et al.  cDNA cloning and sequencing of human fibrillarin, a conserved nucleolar protein recognized by autoimmune antisera. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[60]  M. Stewart,et al.  Separate binding sites on nuclear transport factor 2 (NTF2) for GDP-Ran and the phenylalanine-rich repeat regions of nucleoporins p62 and Nsp1p. , 1996, Journal of molecular biology.

[61]  C. Burd,et al.  hnRNP proteins and the biogenesis of mRNA. , 1993, Annual review of biochemistry.

[62]  A. Shevchenko,et al.  Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. , 1996, Analytical chemistry.

[63]  G. Dreyfuss,et al.  Characterization of the major hnRNP proteins from Drosophila melanogaster , 1992, The Journal of cell biology.

[64]  M. Greaves,et al.  A one-step purification of membrane proteins using a high efficiency immunomatrix. , 1982, The Journal of biological chemistry.

[65]  M. Rosbash,et al.  Identification of a novel nuclear pore-associated protein as a functional target of the HIV-1 Rev protein in yeast , 1995, Cell.

[66]  I. Mattaj,et al.  Nucleocytoplasmic transport: the soluble phase. , 1998, Annual review of biochemistry.

[67]  G. Blobel,et al.  Crystallographic Analysis of the Recognition of a Nuclear Localization Signal by the Nuclear Import Factor Karyopherin α , 1998, Cell.

[68]  E. Buhle,et al.  Correlation between structure and mass distribution of the nuclear pore complex and of distinct pore complex components , 1990, The Journal of cell biology.

[69]  A. Podtelejnikov,et al.  Linking genome and proteome by mass spectrometry: large-scale identification of yeast proteins from two dimensional gels. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[70]  G. Blobel,et al.  The human CAN protein, a putative oncogene product associated with myeloid leukemogenesis, is a nuclear pore complex protein that faces the cytoplasm. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[71]  M. Powers,et al.  Nuclear Export Receptors: From Importin to Exportin , 1997, Cell.

[72]  G. Lipowsky,et al.  Identification of a tRNA-specific nuclear export receptor. , 1998, Molecular cell.

[73]  G. Grosveld,et al.  G2 arrest and impaired nucleocytoplasmic transport in mouse embryos lacking the proto‐oncogene CAN/Nup214. , 1996, The EMBO journal.