Primary sequence and heterologous expression of nuclear pore glycoprotein p62.

Starr, Christopher M.; D’Onofrio, Mara; Park, M K; Hanover, John A.

doi:10.1083/jcb.110.6.1861

Cited by 137 publications

(69 citation statements)

References 27 publications

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“…RanGDP did not bind significantly to RanBP2 ( Figure 5A) or to RanBP1 (our unpublished results) under these conditions. We also used the microtiter plate-binding assay to compare the binding affinity of p97 for RanBP2 and for p62, a nucleoporin that does not bind Ran but that also contains FG repeats (Starr et al, 1990). We found that p97 bound both to RanBP2 ( Figure 5B) and to p62 ( Figure 5C) with an apparent K d of 14 nM.…”

Section: Characterization Of the Binding Of P97 And Ran To Ranbp2mentioning

confidence: 99%

RanGTP Targets p97 to RanBP2, a Filamentous Protein Localized at the Cytoplasmic Periphery of the Nuclear Pore Complex

Delphin

Guan

Melchior

et al. 1997

MBoC

135

126

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RanBP2, a protein containing FG repeat motifs and four binding sites for the guanosine triphosphatase Ran, is localized at the cytoplasmic periphery of the nuclear pore complex (NPC) and is believed to play a critical role in nuclear protein import. We purified RanBP2 from rat liver nuclear envelopes and examined its structural and biochemical properties. Electron microscopy showed that RanBP2 forms a flexible filamentous molecule with a length of ∼36 nm, suggesting that it comprises a major portion of the cytoplasmic fibrils implicated in initial binding of import substrates to the NPC. Using in vitro assays, we characterized the ability of RanBP2 to bind p97, a cytosolic factor implicated in the association of the nuclear localization signal receptor with the NPC. We found that RanGTP promotes the binding of p97 to RanBP2, whereas it inhibits the binding of p97 to other FG repeat nucleoporins. These data suggest that RanGTP acts to specifically target p97 to RanBP2, where p97 may support the binding of an nuclear localization signal receptor/substrate complex to RanBP2 in an early step of nuclear import.

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Section: Characterization Of the Binding Of P97 And Ran To Ranbp2mentioning

confidence: 99%

RanGTP Targets p97 to RanBP2, a Filamentous Protein Localized at the Cytoplasmic Periphery of the Nuclear Pore Complex

Delphin

Guan

Melchior

et al. 1997

MBoC

135

126

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“…The list includes p62 [3,29], Nup153 [31], Nup155 [23], CAN/Nup214 [9], Nup98 [21,22], and RanBP/Nup358 [35,36]. These nucleoporins share the same XFXFG or GLFG repetitive sequence motifs, which are considered to be a nucleoporin signature.…”

mentioning

confidence: 99%

Molecular Cloning and Characterization of a Novel Testis-Specific Nucleoporin-Related Gene

Hd²,

Sy³

et al. 1999

Archives of Andrology

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“…Of the estimated 60-100 different polypeptides that comprise the nuclear pore, only a handful have been identified. These include a component of the basket (nup153), proteins of the central region of the pore (p62/nup60, nup58, and nup54), components of the cytoplasmic filaments (nupl80, nup200/CAN, and TPR), and integral membrane pore proteins (gp210 and POM121), among others (Davis and Blobel, 1986;Starr et al, 1990;Cordes et al, 1991;Finlay et al, 1991;Hallberg et al, 1993;Radu et al, 1993;Sukegawa and Blobel, 1993;Wilken et al, 1993;Byrd et al, 1994; see Rout and Wente, 1994, for a review). A family of pore glycoproteins that contain N-acetylglucosamine residues and consequently bind the lectin WGA are among these pore proteins (for reviews, see Hart et al, 1989;Forbes, 1992;Rout and Wente, 1994).…”

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confidence: 99%

Nuclear pore complex assembly studied with a biochemical assay for annulate lamellae formation.

Meier

Miller

Forbes

1995

The Journal of Cell Biology

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Abstract. Formation of the nuclear pore is an intricate process involving membrane fusion and the ordered assembly of up to 1,000 pore proteins. As such, the study of pore assembly is not a simple one. Interestingly, annulate lamellae, a cytoplasmic organelle consisting of stacks of flattened membrane cisternae perforated by numerous pore complexes, have been found to form spontaneously in a reconstitution system derived from Xenopus egg extracts, as determined by electron microscopy (Dabauvalle et al., 1991). In this work, a biochemical assay for annulate lamellae (AL) formation was developed and used to study the mechanism of AL assembly in general and the assembly of individual nucleoporins into pore complexes in particular. Upon incubation of Xenopus egg cytosol and membrane vesicles, the nucleoporins nup58, nup60, nup97, nup153, and nup200 initially present in a disassembled form in the cytosol became associated with membranes and were pelletable. The association was time and temperature dependent and could be measured by immunoblotting. Thin-section electron microscopy as well as negative staining confirmed that annulate lamellae were forming coincident with the incorporation of pore proteins into membranes. Homogenization and subsequent flotation of the membrane fraction allowed us to separate a population of dense membranes, containing the integral membrane pore protein gp210 and all other nucleoporins tested, from the bulk of cellular membranes. Electron microscopy indicated that annulate lamellae were enriched in this dense, pore protein-containing fraction. GTP~/S prevented incorporation of the soluble pore proteins into membranes. To address whether AL form in the absence of N-acetylglucosaminylated pore proteins, AL assembly was carried out in WGAsepharose--depleted cytosol. Under these conditions, annulate lamellae formed but were altered in appearance. When the membrane fraction containing this altered AL was homogenized and subjected to flotation, the pore protein-containing membranes still sedimented in a distinct peak but were less dense than control annulate lamellae. T HE nuclear pore is responsible for establishing the distinct nuclear and cytoplasmic compartments of the eukaryotic cell. The pore is a highly selective channel through which macromolecular traffic must pass to enter or exit the nucleus. Although ions and other small molecules freely diffuse through the pore, larger molecules require a specific targeting signal for transit (for reviews, see Goldfarb and Michaud,.1991;Forbes, 1992;Gerace, 1992;Osborne and Silver, 1993;Powers and Forbes, 1994).The nuclear pore complex is a large and elaborate structure of ~120 million daltons and is comprised of N1,000 proteins. Structurally, it appears to consist of three ringsThe first two authors, Eva Meier and Brian R. Miller, contributed equally to the results presented here.Please address all correspondence to Douglass J. Forbes, Department of Biology, University of California at San Diego, La,lolla, CA 92093-0347. Tel.: (619) 534-0555....

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Primary sequence and heterologous expression of nuclear pore glycoprotein p62.

Cited by 137 publications

References 27 publications

RanGTP Targets p97 to RanBP2, a Filamentous Protein Localized at the Cytoplasmic Periphery of the Nuclear Pore Complex

RanGTP Targets p97 to RanBP2, a Filamentous Protein Localized at the Cytoplasmic Periphery of the Nuclear Pore Complex

Molecular Cloning and Characterization of a Novel Testis-Specific Nucleoporin-Related Gene

Nuclear pore complex assembly studied with a biochemical assay for annulate lamellae formation.

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