Structure and Assembly of the Nup84p Complex

Siniossoglou, Symeon; Lutzmann, Malik; Santos‐Rosa, Helena; Leonard, Kevin; Mueller, Shirley M.; Aebi, Ueli; Hurt, Ed

doi:10.1083/jcb.149.1.41

Cited by 166 publications

(178 citation statements)

References 31 publications

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“…Isolation under native conditions and in vitro reconstitution have further revealed the Y-shaped structure of this complex (Siniossoglou et al, 2000;Lutzmann et al, 2002). The vertebrate Nup107-160 complex was so far demonstrated to consist of Nup107 (homologous to ScNup84), Nup133 (homologous to ScNup133), Nup160 (homologous to ScNup120 and thus also referred in human cells as Nup120), Nup96 (homologous to ScNup145-C), Sec13 and Nup85/75 (Fontoura et al, 1999;Belgareh et al, 2001;Vasu et al, 2001;Harel et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

The Entire Nup107-160 Complex, Including Three New Members, Is Targeted as One Entity to Kinetochores in Mitosis

Loïodice

Alves

Rabut

et al. 2004

MBoC

254

240

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In eukaryotes, bidirectional transport of macromolecules between the cytoplasm and the nucleus occurs through elaborate supramolecular structures embedded in the nuclear envelope, the nuclear pore complexes (NPCs). NPCs are composed of multiple copies of ϳ30 different proteins termed nucleoporins, of which several can be biochemically isolated as subcomplexes. One such building block of the NPC, termed the Nup107-160 complex in vertebrates, was so far demonstrated to be composed of six different nucleoporins. Here, we identify three WD (Trp-Asp)-repeat nucleoporins as new members of this complex, two of which, Nup37 and Nup43, are specific to higher eukaryotes. The third new member Seh1 is more loosely associated with the Nup107-160 complex biochemically, but its depletion by RNA interference leads to phenotypes similar to knock down of other constituents of this complex. By combining green fluorescent protein-tagged nucleoporins and specific antibodies, we show that all the constituents of this complex, including Nup37, Nup43, Seh1, and Sec13, are targeted to kinetochores from prophase to anaphase of mitosis. Together, our results indicate that the entire Nup107-160 complex, which comprises nearly one-third of the so-far identified nucleoporins, specifically localizes to kinetochores in mitosis.

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Section: Introductionmentioning

confidence: 99%

The Entire Nup107-160 Complex, Including Three New Members, Is Targeted as One Entity to Kinetochores in Mitosis

Loïodice

Alves

Rabut

et al. 2004

MBoC

254

240

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“…The gene encoding the eponymous subunit of the Nup84 NPC subcomplex (NUP84) was independently identified by SGA analysis with a ⌬gcr2 query strain. Deletion of NUP120 or NUP133, which encode two other components of the Nup84 subcomplex (20,21), also caused a synthetic defect in combination with the ⌬gcr1 mutation. Disruption of NUP84, NUP120, or NUP133 is known to cause defects in nuclear membrane and NPC organization (21) and, like the ⌬gcr1 and ⌬gcr2 mutations (16), results in defective growth on glucose.…”

Section: A Synthetic Genetic Interaction Network Suggests a Functionamentioning

confidence: 99%

“…For example, synthetic genetic array (SGA) analysis identified a robust genetic network that connects the Rap1 coactivators Gcr1 and Gcr2 with the Nup84 subcomplex (20,21) and other nuclear envelope components; some of these same perinuclear factors coimmunoprecipitated with Gcr1 and Gcr2.…”

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confidence: 99%

Reverse recruitment: The Nup84 nuclear pore subcomplex mediates Rap1/Gcr1/Gcr2 transcriptional activation

Menon

Sarma

Pasula

et al. 2005

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

123

121

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The recruitment model for gene activation presumes that DNA is a platform on which the requisite components of the transcriptional machinery are assembled. In contrast to this idea, we show here that Rap1͞Gcr1͞Gcr2 transcriptional activation in yeast cells occurs through a large anchored protein platform, the Nup84 nuclear pore subcomplex. Surprisingly, Nup84 and associated subcomplex components activate transcription themselves in vivo when fused to a heterologous DNA-binding domain. The Rap1 coactivators Gcr1 and Gcr2 form an important bridge between the yeast nuclear pore complex and the transcriptional machinery. Nucleoporin activation may be a widespread eukaryotic phenomenon, because it was first detected as a consequence of oncogenic rearrangements in acute myeloid leukemia and related syndromes in humans. These chromosomal translocations fuse a homeobox DNA-binding domain to the human homolog (hNup98) of a transcriptionally active component of the yeast Nup84 subcomplex. We conclude that Rap1 target genes are activated by moving to contact compartmentalized nuclear assemblages, rather than through recruitment of the requisite factors to chromatin by means of diffusion. We term this previously undescribed mechanism ''reverse recruitment'' and discuss the possibility that it is a central feature of eukaryotic gene regulation. Reverse recruitment stipulates that activators work by bringing the DNA to an nuclear pore complex-tethered platform of assembled transcriptional machine components.chromatin boundaries ͉ leukemia ͉ silencing ͉ synthetic genetic array ͉ gene regulation A n underlying assumption of both the stepwise and preassembly alternatives (1) of the recruitment model of in vivo gene activation (2-6) is that activators work by bringing the transcriptional machinery to the DNA, i.e., that the machinery itself diffuses relatively freely within the nuclear compartment. We have been studying the repressor͞activator protein Rap1 of Saccharomyces cerevisiae, which recognizes identical motifs in mediating either transcriptional activation (of glycolytic genes and ribosomal protein genes; refs. 7-9) or repression (of silent mating type loci and telomeres; refs. 10-15) and with its coactivators Gcr1 and Gcr2 participates in coordination of growth with cell-cycle progression (16,17). Numerous aspects of Rap1 activation have conformed poorly with the ''free diffusion'' aspect of the recruitment model for transcriptional activation. One such aspect is the presence of an unusually large activation domain that is easily inactivated by means of mutations throughout the N-terminal 280 residues of Gcr1, spanning four distinct hypomutable regions (8,17,18); two of these hypomutable regions overlap with putative transmembrane domains.We report here independent approaches demonstrating that the Rap1͞Gcr1͞Gcr2 activation assemblage (7-9, 19), like its silencing counterpart, is anchored at the nuclear periphery. For example, synthetic genetic array (SGA) analysis identified a robust genetic network that connects the Ra...

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“…Similar subcomplexes also were obtained by dissecting intact NPCs, using nonionic detergents and a range of salt concentrations (12). In yeast, a well-characterized heptameric subcomplex consists of Nup84, Nup85, Nup120, Nup133, Nup145C, Sec13, and Seh1 (18)(19)(20). Negative-stain electron microscopy of this heptamer assembled from recombinant proteins has revealed a Ϸ400-Å-long, Y-shaped assembly and established the relative position of its members (19).…”

mentioning

confidence: 99%

Structural and functional analysis of Nup120 suggests ring formation of the Nup84 complex

Seo

Debler

et al. 2009

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

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The Nup84 complex constitutes a key building block in the nuclear pore complex (NPC). Here we present the crystal structure of one of its 7 components, Nup120, which reveals a ␤ propeller and an ␣-helical domain representing a novel fold. We discovered a previously unidentified interaction of Nup120 with Nup133 and confirmed the physiological relevance in vivo. As mapping of the individual components in the Nup84 complex places Nup120 and Nup133 at opposite ends of the heptamer, our findings indicate a head-to-tail arrangement of elongated Nup84 complexes into a ring structure, consistent with a fence-like coat for the nuclear pore membrane. The attachment site for Nup133 lies at the very end of an extended unstructured region, which allows for flexibility in the diameter of the Nup84 complex ring. These results illuminate important roles of terminal unstructured segments in nucleoporins for the architecture, function, and assembly of the NPC.crystal structure ͉ fluorescence localization ͉ mRNA export ͉ nuclear pore complex ͉ site-directed mutagenesis

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Structure and Assembly of the Nup84p Complex

Cited by 166 publications

References 31 publications

The Entire Nup107-160 Complex, Including Three New Members, Is Targeted as One Entity to Kinetochores in Mitosis

The Entire Nup107-160 Complex, Including Three New Members, Is Targeted as One Entity to Kinetochores in Mitosis

Reverse recruitment: The Nup84 nuclear pore subcomplex mediates Rap1/Gcr1/Gcr2 transcriptional activation

Structural and functional analysis of Nup120 suggests ring formation of the Nup84 complex

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