Jul 10, 2007 the results indicate that the maximal size of protein to diffuse through the nuclear pore complex could be quite larger than 60kda, thus greatly extending the diffusion limit that the nuclear pore can accommodate. Nuclear pore complexes npcs fuse the inner and outer nuclear membranes to form channels across the nuclear envelope. Extremely longlived nuclear pore proteins in the rat brain. Mar, 2002 proteomic studies have the potential to comprehensively define the composition of organelles but are limited by the organellar crosscontamination that arises during subcellular fractionation. To further understand the structure and function of the mammalian npc, we have completed a proteomic analysis to identify and classify all of its. These protein interactions can occur on radically different time scales. The turnover of each protein in the mammalian proteome is a functionally important characteristic. Jan 01, 2006 schematic diagram of the nuclear pore complex located within the nuclear envelope ne at the interface of cytoplasm c and nucleus n indicating the approximate location of the nucleoporins and nft2 immunolabeled in this study.
Structure and evolution of mammalian ribosomal proteins. Npc is a large protein structure, spanning the double nuclear membrane and enabling nucleocytoplasmic exchange 11. An understanding of how the nuclear pore complex npc mediates nucleocytoplasmic exchange requires a comprehensive inventory of the molecular components of the npc and a knowledge of how each. The structure of the nuclear pore complex nuclear pore complexes are proteinaceous structures embedded in the double membrane of the nuclear envelope. The npc is essential for the transport of biomolecules across the nuclear envelope. In animal cells, involvement of the nuclear pore complex in morphology. Its octagonal ring scaffold perforates the nuclear envelope and features a unique molecular machinery that regulates nucleocytoplasmic transport. The former contains 16 s and the latter contains 12 s rrnas as rna components. Matunis 1 1 department of biochemistry and molecular biology, bloomberg school of public health, johns hopkins university, baltimore, md 21205 2. Cronshaw 1 department of biochemistry and molecular biology, bloomberg school of public health, johns hopkins university, baltimore, md 21205. It is a double membrane system continuous with the endoplasmic reticulum er that consists of three connected but distinguishable membrane domains. Nuclear pore complexes npcs are indispensable for cell function and are at the center of several human diseases.
These ellps were associated with chromatin and the nuclear pore complex, the central transport channels. Here, we explore the domain structure of nup3, a nucleoporin in a conserved npc subcomplex that is crucial for npc biogenesis and is believed to form part of the npc scaffold. As the sole site of nucleocytoplasmic transport, the nuclear pore complex npc has a vital cellular role. They are large macromolecular assemblies with a complex composition and diverse functions. Kentaro tamura, ikuko haranishimura, the molecular architecture of the plant nuclear pore complex. The molecular architecture of the nuclear pore complex. Apr 24, 2018 the nuclear pore complex npc is one of the largest protein assemblies in eukaryotes comprising over 500 nucleoporin subunits. Some proteins form relatively permanent associations with the core structure, and so are termed nuclear pore complex components or nucleoporins nups. Although the npc is a complex structure, our analysis reveals underlying simplicities in its architecture. Many identified rhythmic proteins were parts of nuclear complexes involved in transcriptional regulation, ribosome biogenesis, dna repair, and the cell cycle and its potentially associated diurnal rhythm of hepatocyte polyploidy. Nonetheless, much remains to be learned about many.
Aug 17, 2010 the nuclear pore complex npc is the sole mediator of transport between the nucleus and the cytoplasm. Proteomic analysis of the mammalian nuclear pore complex janet m. The npc is composed of about 30 distinct proteins, termed nucleoporins or nups. Proteomic analysis of the mammalian mitochondrial ribosome. Nucleoporin 37 nup37 is a protein that in humans is encoded by the nup37 gene function. Superresolution mapping of scaffold nucleoporins in the. The nuclear pore complex npc is a macromolecular assembly embedded within the nuclear envelope that mediates. Direct involvement of the inner nuclear envelope protein in npc anchoring and positioning was suggested by studies in vertebrate cells. A proteomic analysis tianwei li, evgenij evdokimov, rongfong shen, chienchung chao, ephrem tekle, tao wang. Mca39c7 was one of a series of clones which strongly and specifically labelled the nuclear pore complex.
Nuclear proteomics uncovers diurnal regulatory landscapes. Thirteen nucleoporins that contain fg peptide repeats fg nups are proposed to function as stepping stones in karyopherinmediated transport pathways. Stripping the megadalton complex down to its most essential organizational elements, one can divide the npc into scaffold components and the disordered elements attached to it that generate a selective barrier between compartments. Contribution of electron microscopy to the study of the. When this antibody was tested on cells from other species, including rat, mouse and human cells, it has invariably strongly stained nuclear pore complexes, so it appears to be an excellent and panspecific marker for these important structures. Structural and functional analysis of nup3 domains reveals. Report comparative genomics, evolution and origins of the. It is a large supramolecular complex made up of multiple. The fold composition of the nucleoporins forming the scaffold is remarkably simple. Structural analysis of the p62 complex, an assembly of olinked glycoproteins that localizes near the central gated channel of the nuclear pore complex. The components of the mammalian npc were identified and proteomically analyzed in. Feb 24, 2012 to combat the functional decline of the proteome, cells use the process of protein turnover to replace potentially impaired polypeptides with new functional copies. Nuclear pore complexes npcs perforate the nuclear envelope and serve as the primary transport gates for molecular exchange between nucleus and cytoplasm. The nuclear pore complex as a flexible and dynamic gate.
The molecular architecture of the nuclear pore complex eriba. Cronshaw jm1, krutchinsky an, zhang w, chait bt, matunis mj. Stoichiometry and compositional plasticity of the yeast. Here, we employed highresolution mass spectrometry to quantify protein dynamics in nondividing mammalian cells. Crystal structure of a human nuclear pore complex component nup43.
The nuclear pore complex npc is responsible for nucleocytoplasmic transport and constitutes a hub for control of gene expression. Apart from facilitating nucleocytoplasmic transport, npcs are involved in chromatin organization. The nuclear pore complex structure and function at a glance. The permeability barrier of nuclear pore complexes npcs controls the exchange between nucleus and cytoplasm. Aladin mutations lead to mistargeting of the protein in the npc. Pan specific nuclear pore complex mouse monoclonal antibody. They are aqueous channels generated from a complex network of evolutionarily conserved proteins known as nucleporins. 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. At its heart, the npc contains a highly connected scaffold that attaches to and coats the curved pore membrane. The nuclear pore complex npc is the sole gateway between the nucleus and the cytoplasm of interphase eukaryotic cells, and it mediates all trafficking between these 2 cellular compartments. Cronshaw jm, krutchinsky an, zhang w et al 2002 proteomic analysis of the mammalian nuclear pore complex. They are large macromolecular assemblies with a complex composition and. Interactome mapping reveals the evolutionary history of the. Large macromolecules are actively transported through the nuclear pore complex through the central transporter complex.
Nuclear pore complex proteins in alzheimer disease journal. Nuclear pore complexes in the organization and regulation of. Nuclear proteomics uncovers diurnal regulatory landscapes in. Npc structures of amazingly similar appearance have been identified in all eukaryotes from yeast to human, yet differences in npcs between species are likely to provide important clues to npc function.
Npcs provide access to the nucleus and regulate the transport of proteins and rna across the nuclear envelope. Comparative proteomics of organellar subfractions can mitigate these problems, as demonstrated by a recent study involving the nuclear envelope. We show here that a saturated hydrogel formed by a single nucleoporin fgrepeat domain is sufficient to. Citeseerx document details isaac councill, lee giles, pradeep teregowda. An understanding of how the nuclear pore complex npc mediates nucleocytoplasmic exchange requires a comprehensive inventory of the molecular components of the npc and a. Systemswide proteomic analysis in mammalian cells reveals conserved, functional protein turnover. Jcb article proteomic analysis of the mammalian nuclear pore. Matunis 1 1 department of biochemistry and molecular biology, bloomberg school of public health, johns hopkins university, baltimore, md 21205. Parallel analysis of the nuclear phosphoproteome enabled the inference of the temporal activity of kinases accounting for rhythmic phosphorylation. Involvement of the nuclear pore complex in morphology of the.
The nuclear pore complex npc, which spans two nuclear membranes, regulates the exchange of macromolecule trafficking between the nucleus and the cytoplasm. The molecular architecture of the nuclear pore complex nature. The maximal size of protein to diffuse through the nuclear. The saccharomyces cerevisiae nuclear pore complex is a supramolecular assembly of 30 nucleoporins that cooperatively facilitate nucleocytoplasmic transport. Comparative proteomic analyses of the nuclear envelope and. Proteomic analysis of nucleoporin interacting proteins journal of. Transport of macromolecules between the cytoplasm and nucleus occurs through nuclear pore complexes npcs embedded in the nuclear envelope.
Using subsaturating silac labeling, we obtained accurate turnover rates of. Aug 31, 2001 the nuclear pore complex npc forms the conduit for the exchange of information between the nucleus and cytoplasm. Nov 29, 2007 the nuclear pore complex plays a crucial role in the cell, as gatekeeper for traffic between the cytoplasm and the interior of the nucleus. Nonetheless, much remains to be learned about many fundamental aspects of npc function. The components of npcs from several eukaryotic lineages have been determined, but only the yeast and vertebrate npcs have been extensively characterized at the quaternary level. Proteomic analysis of nucleoporin interacting proteins. Evidence for a shared nuclear pore complex architecture that is. We found that extremely longlived proteins ellps did not turn over in postmitotic cells of the rat central nervous system. In addition, immunogold electron microscopy has been used to reveal the molecular composition of the nuclear pore complex, and to dissect nuclear transport into distinct steps. Aladin is one of the nucleoporins constituting the nuclear pore complex npc and belongs to the wdrepeat protein family. Sumoylation of heterogeneous nuclear ribonucleoproteins, zinc. Proteomic analysis of the mammalian nuclear pore complex.
In order to understand how the npc functions, it is useful to examine the similarities and differences between npcs from the yeast saccharomyces cerevisiae and the frog xenopus laevis. Proteomic analysis of the mammalian nuclear pore complex article in the journal of cell biology 1585. While the human and mouse genomes have been available for over a decade, progress in measuring the expression of gene products has been made mainly on the level of mrna abundance mele et al. However, the precise copy number and the spatial location of each nup in the native npc remain. Herein, we have developed a widely applicable imaging pipeline to determine the. The mammalian mitochondrial ribosome mitoribosome 1 has a smaller sedimentation coefficient 55 s, compared with the bacterial ribosome 70 s, and consists of large 39 s and small 28 s subunits. Dec 21, 2016 nuclear pore complexes npcs fuse the inner and outer nuclear membranes to form channels across the nuclear envelope. The nuclear pore complex plays a crucial role in the cell, as gatekeeper for traffic between the cytoplasm and the interior of the nucleus. Nucleocytoplasmic transport is mediated by nuclear pore complexes npcs allen et al. Sep 02, 2002 proteomic analysis of the mammalian nuclear pore complex janet m. Nov 22, 2004 nucleocytoplasmic transport occurs through nuclear pore complexes npcs whose complex architecture is generated from a set of only. Sumoylation of heterogeneous nuclear ribonucleoproteins, zinc finger proteins, and nuclear pore complex proteins.