Proteomic analysis of the mammalian nuclear pore complex

As the sole site of nucleocytoplasmic transport, the nuclear pore complex (NPC) has a vital cellular role. Nonetheless, much remains to be learned about many fundamental aspects of NPC function. To further understand the structure and function of the mammalian NPC, we have completed a proteomic analysis to identify and classify all of its protein components. We used mass spectrometry to identify all proteins present in a biochemically purified NPC fraction. Based on previous characterization, sequence homology, and subcellular localization, 29 of these proteins were classified as nucleoporins, and a further 18 were classified as NPC-associated proteins. Among the 29 nucleoporins were six previously undiscovered nucleoporins and a novel family of WD repeat nucleoporins. One of these WD repeat nucleoporins is ALADIN, the gene mutated in triple-A (or Allgrove) syndrome. Our analysis defines the proteome of the mammalian NPC for the first time and paves the way for a more detailed characterization of NPC structure and function.

[1]  H. Saitoh,et al.  Enzymes of the SUMO Modification Pathway Localize to Filaments of the Nuclear Pore Complex , 2002, Molecular and Cellular Biology.

[2]  M. Dasso,et al.  Association of the Human SUMO-1 Protease SENP2 with the Nuclear Pore* , 2002, The Journal of Biological Chemistry.

[3]  U. K. Laemmli,et al.  Chromatin Boundaries in Budding Yeast The Nuclear Pore Connection , 2002, Cell.

[4]  D. Lacombe,et al.  Progressive bulbospinal amyotrophy in triple A syndrome with AAAS gene mutation. , 2002, Neurology.

[5]  R. Orrell,et al.  ALADIN, but where's the genie? , 2002, Neurology.

[6]  Ed Hurt,et al.  A Conserved mRNA Export Machinery Coupled to pre-mRNA Splicing , 2002, Cell.

[7]  A. Dejean,et al.  The Nucleoporin RanBP2 Has SUMO1 E3 Ligase Activity , 2002, Cell.

[8]  A. Huebner,et al.  Clinical and Novel Molecular Findings in a 6.8-Year-Old Turkish Boy with Triple A Syndrome , 2002, Hormone Research in Paediatrics.

[9]  R. Thakker.  Multiple Endocrine Neoplasia , 2001, Hormone Research in Paediatrics.

[10]  I. Macara.  Transport into and out of the Nucleus , 2001, Microbiology and Molecular Biology Reviews.

[11]  C. Stratakis,et al.  Spectrum of mutations of the AAAS gene in Allgrove syndrome: lack of mutations in six kindreds with isolated resistance to corticotropin. , 2001, The Journal of clinical endocrinology and metabolism.

[12]  W. Fischer,et al.  Novel vertebrate nucleoporins Nup133 and Nup160 play a role in mRNA export , 2001, The Journal of cell biology.

[13]  B. Chait,et al.  Automatic identification of proteins with a MALDI-quadrupole ion trap mass spectrometer. , 2001, Analytical chemistry.

[14]  N. Daigle,et al.  An evolutionarily conserved NPC subcomplex, which redistributes in part to kinetochores in mammalian cells , 2001, The Journal of cell biology.

[15]  W. Chin,et al.  Identification and Characterization of RRM-containing Coactivator Activator (CoAA) as TRBP-interacting Protein, and Its Splice Variant as a Coactivator Modulator (CoAM)* , 2001, The Journal of Biological Chemistry.

[16]  S. Vasu,et al.  Nuclear pores and nuclear assembly. , 2001, Current opinion in cell biology.

[17]  J. Aitchison,et al.  A Link between the Synthesis of Nucleoporins and the Biogenesis of the Nuclear Envelope , 2001, The Journal of cell biology.

[18]  K Ribbeck,et al.  Kinetic analysis of translocation through nuclear pore complexes , 2001, The EMBO journal.

[19]  J. Hauber,et al.  Cofactor Requirements for Nuclear Export of Rev Response Element (Rre)–And Constitutive Transport Element (Cte)–Containing Retroviral Rnas , 2001, The Journal of cell biology.

[20]  B. Graveley.  Alternative splicing: increasing diversity in the proteomic world. , 2001, Trends in genetics : TIG.

[21]  S. Hennig,et al.  Triple A syndrome is caused by mutations in AAAS, a new WD-repeat protein gene. , 2001, Human molecular genetics.

[22]  Catherine G. Takizawa,et al.  Control of mitosis by changes in the subcellular location of cyclin-B1-Cdk1 and Cdc25C. , 2000, Current opinion in cell biology.

[23]  Claude Mugnier,et al.  Mutant WD-repeat protein in triple-A syndrome , 2000, Nature Genetics.

[24]  G. Blobel,et al.  The Nucleoporin Nup98 Is a Site for GDP/GTP Exchange on Ran and Termination of Karyopherin β2-mediated Nuclear Import* , 2000, The Journal of Biological Chemistry.

[25]  C. Smythe,et al.  Incorporation of the nuclear pore basket protein Nup153 into nuclear pore structures is dependent upon lamina assembly: evidence from cell‐free extracts of Xenopus eggs , 2000, The EMBO journal.

[26]  B. Chait,et al.  Rapidly switchable matrix-assisted laser desorption/ionization and electrospray quadrupole-time-of-flight mass spectrometry for protein identification , 2000, Journal of the American Society for Mass Spectrometry.

[27]  A. Capobianco,et al.  Neoplastic Transformation by Notch Requires Nuclear Localization , 2000, Molecular and Cellular Biology.

[28]  S. R. Wente,et al.  The nuclear pore complex: a protein machine bridging the nucleus and cytoplasm. , 2000, Current opinion in cell biology.

[29]  R. Dohmen,et al.  SUMO conjugation and deconjugation , 2000, Molecular and General Genetics MGG.

[30]  E. Kiseleva,et al.  The nuclear pore complex: mediator of translocation between nucleus and cytoplasm. , 2000, Journal of cell science.

[31]  B. Chait,et al.  ProFound: an expert system for protein identification using mass spectrometric peptide mapping information. , 2000, Analytical chemistry.

[32]  B. Chait,et al.  The Yeast Nuclear Pore Complex: Composition, Architecture, and Transport Mechanism , 2000 .

[33]  Jean-Christophe Olivo-Marin,et al.  Nuclear pore complexes in the organization of silent telomeric chromatin , 2000, Nature.

[34]  D. Lane,et al.  Accumulating active p53 in the nucleus by inhibition of nuclear export: a novel strategy to promote the p53 tumor suppressor function. , 1999, Experimental cell research.

[35]  D. N. Perkins,et al.  Probability‐based protein identification by searching sequence databases using mass spectrometry data , 1999, Electrophoresis.

[36]  T. Lange,et al.  Cell cycle dependent localization of the telomeric PARP, tankyrase, to nuclear pore complexes and centrosomes. , 1999, Journal of cell science.

[37]  G. Blobel,et al.  GTP Hydrolysis Links Initiation and Termination of Nuclear Import on the Nucleoporin Nup358* , 1999, The Journal of Biological Chemistry.

[38]  J. Hanover,et al.  Phosphorylation and glycosylation of nucleoporins. , 1999, Archives of biochemistry and biophysics.

[39]  G. Blobel,et al.  Putative Reaction Intermediates in Crm1-mediated Nuclear Protein Export* , 1999, The Journal of Biological Chemistry.

[40]  Temple F. Smith,et al.  The WD repeat: a common architecture for diverse functions. , 1999, Trends in biochemical sciences.

[41]  G. Dreyfuss,et al.  Nup153 is an M9‐containing mobile nucleoporin with a novel Ran‐binding domain , 1999, The EMBO journal.

[42]  B. Fontoura,et al.  A Conserved Biogenesis Pathway for Nucleoporins: Proteolytic Processing of a 186-Kilodalton Precursor Generates Nup98 and the Novel Nucleoporin, Nup96 , 1999, The Journal of cell biology.

[43]  J. Aitchison,et al.  Specific Binding of the Karyopherin Kap121p to a Subunit of the Nuclear Pore Complex Containing Nup53p, Nup59p, and Nup170p , 1998, The Journal of cell biology.

[44]  T. Giddings,et al.  Saccharomyces cerevisiae Ndc1p Is a Shared Component of Nuclear Pore Complexes and Spindle Pole Bodies , 1998, The Journal of cell biology.

[45]  L. Gerace,et al.  cDNA cloning and analysis of the expression of nucleoporin p45. , 1998, Gene.

[46]  J. L. Watkins,et al.  The human homologue of Saccharomyces cerevisiae Gle1p is required for poly(A)+ RNA export. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[47]  B. Chait,et al.  Protein indentification using mass spectrometric information , 1998, Electrophoresis.

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

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

[50]  M. Bornens,et al.  Identification of a new mammalian centrin gene, more closely related to Saccharomyces cerevisiae CDC31 gene. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[51]  Zhang Jz,et al.  Immunogold labeling electron microscopy showing vimentin filament anchored on nuclear pore complex , 1997 .

[52]  M. Dasso,et al.  RanBP2 associates with Ubc9p and a modified form of RanGAP1. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[53]  F. Melchior,et al.  A Small Ubiquitin-Related Polypeptide Involved in Targeting RanGAP1 to Nuclear Pore Complex Protein RanBP2 , 1997, Cell.

[54]  S. Cai,et al.  [Immunogold labeling electron microscopy showing vimentin filament anchored on nuclear pore complex]. , 1997, Shi yan sheng wu xue bao.

[55]  G. Blobel,et al.  A novel ubiquitin-like modification modulates the partitioning of the Ran-GTPase-activating protein RanGAP1 between the cytosol and the nuclear pore complex , 1996, The Journal of cell biology.

[56]  F. Jeanmougin,et al.  A possible involvement of TIF1 alpha and TIF1 beta in the epigenetic control of transcription by nuclear receptors. , 1996, The EMBO journal.

[57]  R. Wozniak,et al.  Cell cycle-dependent phosphorylation of nucleoporins and nuclear pore membrane protein Gp210. , 1996, Biochemistry.

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

[59]  M. Dabauvalle,et al.  Localization of the Ran-GTP binding protein RanBP2 at the cytoplasmic side of the nuclear pore complex. , 1995, European journal of cell biology.

[60]  S. Müller,et al.  Structural analysis of the p62 complex, an assembly of O-linked glycoproteins that localizes near the central gated channel of the nuclear pore complex. , 1995, Molecular biology of the cell.

[61]  I. Wool,et al.  Structure and evolution of mammalian ribosomal proteins. , 1995, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[62]  Tatsuya Seki,et al.  A giant nucleopore protein that binds Ran/TC4 , 1995, Nature.

[63]  G. Blobel,et al.  Nup358, a Cytoplasmically Exposed Nucleoporin with Peptide Repeats, Ran-GTP Binding Sites, Zinc Fingers, a Cyclophilin A Homologous Domain, and a Leucine-rich Region (*) , 1995, The Journal of Biological Chemistry.

[64]  L. Orci,et al.  Human SEC13Rp functions in yeast and is located on transport vesicles budding from the endoplasmic reticulum , 1995, The Journal of cell biology.

[65]  F. Masiarz,et al.  Reconstituted nuclei depleted of a vertebrate GLFG nuclear pore protein, p97, import but are defective in nuclear growth and replication , 1995, The Journal of cell biology.

[66]  D. Forbes,et al.  Differential Mitotic Phosphorylation of Proteins of the Nuclear Pore Complex (*) , 1995, The Journal of Biological Chemistry.

[67]  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.

[68]  M. Rout,et al.  Isolation of the yeast nuclear pore complex , 1993, The Journal of cell biology.

[69]  G. Blobel,et al.  An integral membrane protein of the pore membrane domain of the nuclear envelope contains a nucleoporin-like region , 1993, The Journal of cell biology.

[70]  M. Radermacher,et al.  Architecture of the Xenopus nuclear pore complex revealed by three- dimensional cryo-electron microscopy , 1993, The Journal of cell biology.

[71]  G. Blobel,et al.  Identification of a receptor for protein import into mitochondria , 1990, Nature.

[72]  U. Greber,et al.  A major glycoprotein of the nuclear pore complex is a membrane‐spanning polypeptide with a large lumenal domain and a small cytoplasmic tail. , 1990, The EMBO journal.

[73]  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.

[74]  T. Gerken,et al.  Role of glycosylation on the conformation and chain dimensions of O-linked glycoproteins: light-scattering studies of ovine submaxillary mucin. , 1989, Biochemistry.

[75]  G. Blobel,et al.  Nuclear pore complex contains a family of glycoproteins that includes p62: glycosylation through a previously unidentified cellular pathway. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[76]  D. Newmeyer,et al.  Inhibition of in vitro nuclear transport by a lectin that binds to nuclear pores , 1987, The Journal of cell biology.

[77]  G. Blobel,et al.  Identification and characterization of a nuclear pore complex protein , 1986, Cell.

[78]  G. Blobel,et al.  A modified procedure for the isolation of a pore complex-lamina fraction from rat liver nuclei , 1976, The Journal of cell biology.

[79]  V. Potter,et al.  Nuclei from Rat Liver: Isolation Method That Combines Purity with High Yield , 1966, Science.