The Three-Dimensional Structure of the Autoproteolytic, Nuclear Pore-Targeting Domain of the Human Nucleoporin Nup98

Hodel, A.E.; Hodel, Mary R.; Griffis, Eric R.; Hennig, Krista A; Ratner, Gary A.; Xu, Shan; Powers, Maureen A.

doi:10.1016/s1097-2765(02)00589-0

Cited by 119 publications

(164 citation statements)

References 32 publications

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“…The Nup98 polypeptide chain encodes one small structured domain at its C terminus, which possesses autoproteolytic activity and thereby facilitates the evolutionarily conserved, cotranslational cleavage of the Nup98 and Nup98-Nup96 precursor proteins (21)(22)(23) (Fig. 1A and Fig.…”

Section: Resultsmentioning

confidence: 99%

Structural and functional analysis of the interaction between the nucleoporin Nup98 and the mRNA export factor Rae1

Ren

Seo

Blobel

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

104

134

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The export of mRNAs is a multistep process, involving the packaging of mRNAs into messenger ribonucleoprotein particles (mRNPs), their transport through nuclear pore complexes, and mRNP remodeling events prior to translation. Ribonucleic acid export 1 (Rae1) and Nup98 are evolutionarily conserved mRNA export factors that are targeted by the vesicular stomatitis virus matrix protein to inhibit host cell nuclear export. Here, we present the crystal structure of human Rae1 in complex with the Gle2-binding sequence (GLEBS) of Nup98 at 1.65 Å resolution. Rae1 forms a seven-bladed β-propeller with several extensive surface loops. The Nup98 GLEBS motif forms an ≈50-Å-long hairpin that binds with its C-terminal arm to an essentially invariant hydrophobic surface that extends over the entire top face of the Rae1 β-propeller. The C-terminal arm of the GLEBS hairpin is necessary and sufficient for Rae1 binding, and we identify a tandem glutamate element in this arm as critical for complex formation. The Rae1•Nup98 GLEBS surface features an additional conserved patch with a positive electrostatic potential, and we demonstrate that the complex possesses single-stranded RNAbinding capability. Together, these data suggest that the Rae1•Nup98 complex directly binds to the mRNP at several stages of the mRNA export pathway.mRNA export machinery | fluorescence localization | microscopy | site-directed mutagenesis | protein-protein interaction

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Section: Resultsmentioning

confidence: 99%

Structural and functional analysis of the interaction between the nucleoporin Nup98 and the mRNA export factor Rae1

Ren

Seo

Blobel

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

104

134

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“…22 Reminiscent of this chaperoning function, we have shown here that PIDD requires Hsp90 for an auto-processing competent conformation. Similar to PIDD, the nucleoporin Nup98 also exhibits strict structural requirements for HFS site auto-cleavage to occur; 29,30 it would be interesting to establish whether Nup98 requires chaperones for proteolytic activity analogous to PIDD. The structural details of this Hsp90-dependent conformation remain to be investigated, but the need for chaperones appears to be common among multi-domain proteins, which have to balance structural confinements with function.…”

Section: Discussionmentioning

confidence: 99%

Regulation of PIDD auto-proteolysis and activity by the molecular chaperone Hsp90

et al. 2010

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In response to DNA damage, p53-induced protein with a death domain (PIDD) forms a complex called the PIDDosome, which either consists of PIDD, RIP-associated protein with a death domain and caspase-2, forming a platform for the activation of caspase-2, or contains PIDD, RIP1 and NEMO, important for NF-jB activation. PIDDosome activation is dependent on autoprocessing of PIDD at two different sites, generating the fragments PIDD-C and PIDD-CC. Despite constitutive cleavage, endogenous PIDD remains inactive. In this study, we screened for novel PIDD regulators and identified heat shock protein 90 (Hsp90) as a major effector in both PIDD protein maturation and activation. Hsp90, together with p23, binds PIDD and inhibition of Hsp90 activity with geldanamycin efficiently disrupts this association and impairs PIDD auto-processing. Consequently, both PIDD-mediated NF-jB and caspase-2 activation are abrogated. Interestingly, PIDDosome formation itself is associated with Hsp90 release. Characterisation of cytoplasmic and nuclear pools of PIDD showed that active PIDD accumulates in the nucleus and that only cytoplasmic PIDD is bound to Hsp90. Finally, heat shock induces Hsp90 release from PIDD and PIDD nuclear translocation. Thus, Hsp90 has a major role in controlling PIDD functional activity.

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“…It is easily identifiable as it encoded in most eukaryotic genomes as a single protein comprised of a large N-terminal FG-Nup domain and a large C-terminal a-solenoid separated by a b-sandwich that contains within it catalytic residues that lead to its autoproteolytic cleavage into 2 separate proteins. 23 However, although the 2 proteins still associate with one another in the NPC, it is generally believed that cleavage into 2 different polypeptides allows the FG-Nup domain of this protein to be dynamic and have a role in gene regulation, while the a-solenoid portion remains a stable structural member of the outer ring complex. 24,25 The tripeptide catalytic residues in the trypanosome ortholog (TbNup158) have altered during evolution to prevent cleavage, resulting in the protein being incorporated into the TbNPC outer ring as a single protein, possibly to eliminate this dynamism and negate a role in gene regulation which is unusual in trypanosomes (see below and Fig.…”

Section: Copy and Paste: The Npc's Scaffold Arose Through Ancient Dupmentioning

confidence: 99%

Comparative interactomics provides evidence for functional specialization of the nuclear pore complex

Obado

Field

Rout

2017

Nucleus

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The core architecture of the eukaryotic cell was established well over one billion years ago, and is largely retained in all extant lineages. However, eukaryotic cells also possess lineagespecific features, frequently keyed to specific functional requirements. One quintessential core eukaryotic structure is the nuclear pore complex (NPC), responsible for regulating exchange of macromolecules between the nucleus and cytoplasm as well as acting as a nuclear organizational hub. NPC architecture has been best documented in one eukaryotic supergroup, the Opisthokonts (e.g. Saccharomyces cerevisiae and Homo sapiens), which although compositionally similar, have significant variations in certain NPC subcomplex structures. The variation of NPC structure across other taxa in the eukaryotic kingdom however, remains poorly understood. We explored trypanosomes, highly divergent organisms, and mapped and assigned their NPC proteins to specific substructures to reveal their NPC architecture. We showed that the NPC central structural scaffold is conserved, likely across all eukaryotes, but more peripheral elements can exhibit very significant lineage-specific losses, duplications or other alterations in their components. Amazingly, trypanosomes lack the major components of the mRNA export platform that are asymmetrically localized within yeast and vertebrate NPCs. Concomitant with this, the trypanosome NPC is ALMOST completely symmetric with the nuclear basket being the only major source of asymmetry. We suggest these features point toward a stepwise evolution of the NPC in which a coating scaffold first stabilized the pore after which selective gating emerged and expanded, leading to the addition of peripheral remodeling machineries on the nucleoplasmic and cytoplasmic sides of the pore.

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The Three-Dimensional Structure of the Autoproteolytic, Nuclear Pore-Targeting Domain of the Human Nucleoporin Nup98

Cited by 119 publications

References 32 publications

Structural and functional analysis of the interaction between the nucleoporin Nup98 and the mRNA export factor Rae1

Structural and functional analysis of the interaction between the nucleoporin Nup98 and the mRNA export factor Rae1

Regulation of PIDD auto-proteolysis and activity by the molecular chaperone Hsp90

Comparative interactomics provides evidence for functional specialization of the nuclear pore complex

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