cDNA Cloning and Characterization ofNpap60:A Novel Rat Nuclear Pore-Associated Protein with an Unusual Subcellular Localization during Male Germ Cell Differentiation

Fan, Fan; Liu, Cheng-Pin; Korobova, Olga; Heyting, C.; Offenberg, H.H.; Trump, Gary N.; Arnheim, Norman

doi:10.1006/geno.1996.4557

Cited by 40 publications

(41 citation statements)

References 24 publications

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“…We targeted Nup60 (hNup153) and Nup53, found primarily on the nuclear basket (Loeb et al 1993;Fan et al 1997;Hood et al 2000), and Nup100, Nup157, and Nup84, which interact with other parts of the NPC (Aitchison and Rout 2012; Figure 1D). Neither nup53D, nup100D, nor nup157D showed a decrease in spore viability, delayed meiotic progression, or other obvious growth defects ( Figure 1E and Figure S2).…”

Section: Identification Of Proteins That Copurify With Ndj1mentioning

confidence: 99%

The Nucleoporin Nup2 Contains a Meiotic-Autonomous Region that Promotes the Dynamic Chromosome Events of Meiosis

Chu

Gromova²,

Newman³

et al. 2017

Genetics

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Meiosis is a specialized cellular program required to create haploid gametes from diploid parent cells. Homologous chromosomes pair, synapse, and recombine in a dynamic environment that accommodates gross chromosome reorganization and significant chromosome motion, which are critical for normal chromosome segregation. In Saccharomyces cerevisiae, Ndj1 is a meiotic telomere-associated protein required for physically attaching telomeres to proteins embedded in the nuclear envelope. In this study, we identified additional proteins that act at the nuclear periphery from meiotic cell extracts, including Nup2, a nonessential nucleoporin with a known role in tethering interstitial chromosomal loci to the nuclear pore complex. We found that deleting NUP2 affects meiotic progression and spore viability, and gives increased levels of recombination intermediates and products. We identified a previously uncharacterized 125 aa region of Nup2 that is necessary and sufficient for its meiotic function, thus behaving as a meiotic autonomous region (MAR). Nup2-MAR forms distinct foci on spread meiotic chromosomes, with a subset overlapping with Ndj1 foci. Localization of Nup2-MAR to meiotic chromosomes does not require Ndj1, nor does Ndj1 localization require Nup2, suggesting these proteins function in different pathways, and their interaction is weak or indirect. Instead, several severe synthetic phenotypes are associated with the nup2D ndj1D double mutant, including delayed turnover of recombination joint molecules, and a failure to undergo nuclear divisions without also arresting the meiotic program. These data suggest Nup2 and Ndj1 support partially overlapping functions that promote two different levels of meiotic chromosome organization necessary to withstand a dynamic stage of the eukaryotic life cycle.

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Section: Identification Of Proteins That Copurify With Ndj1mentioning

confidence: 99%

The Nucleoporin Nup2 Contains a Meiotic-Autonomous Region that Promotes the Dynamic Chromosome Events of Meiosis

Chu

Gromova²,

Newman³

et al. 2017

Genetics

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“…The final wash contained DAPI (4Ј,6Ј-diamidino-2-phenylindole) to visualize cell nuclei. For methanol-acetone fixation, coverslips were fixed in methanol for 30 min at Ϫ20°C and then quickly rinsed in acetone at 4°C as described earlier (6). After the acetone rinse, the coverslips were washed for 5 min in 1ϫ PBS and then treated as described above.…”

Section: Antibodiesmentioning

confidence: 99%

“…Nup50 was initially described as a rat NPC pro-tein that undergoes unusual changes in localization during male germ cell differentiation (6). We have found that Nup50 is a ubiquitously expressed NPC protein that exhibited twohybrid interactions with multiple transport receptor proteins and coimmunoprecipitated with the nucleoporin Nup153.…”

mentioning

confidence: 99%

Characterization and Targeted Disruption of Murine Nup50, a p27^Kip1-Interacting Component of the Nuclear Pore Complex

Smitherman

Lee

Swanger

et al. 2000

Molecular and Cellular Biology

105

104

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p27Kip1 is a member of the Cip-Kip family of cyclin-dependent kinase (Cdk) inhibitors that binds to cyclin-Cdk complexes and inhibits their catalytic activity in response to antiproliferative stimuli. . We found that murine Nup50 is a widely expressed nucleoporin and that Nup50 expression is highest in the developing neural tube and adult testes. We have also examined interactions between Nup50 and the NPC and found specific two-hybrid interactions between Nup50 and several well-defined components of the NPC, as well as coimmunoprecipitation of Nup50 with the nucleoporin Nup153 from transfected mammalian cells. In order to study Nup50 function in vivo, we cloned the mouse Nup50 genomic locus and created a targeted Nup50 deletion in the mouse germ line. Nup50 disruption resulted in a complex phenotype characterized by late embryonic lethality, neural tube defects, and intrauterine growth retardation. Although Nup50-null mouse embryo fibroblasts exhibited no defects in either cell cycle control or p27 Kip1 regulation, Nup50 deletion was associated with abnormalities in p27 Kip1 expression and cell proliferation in the developing neuroepithelium. We conclude that Nup50 is a nucleoporin with essential functions during mouse development.

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“…NPC proteins (nucleoporins) located at the nuclear side of the vertebrate NPC comprise Nup153 (Sukegawa and Blobel, 1993), Nup98 (Powers et al, 1995;Radu et al, 1995), Nup 50 (Fan et al, 1997;Guan et al, 2000;Smitherman et al, 2000), the components of the Nup160-Nup133-Nup96-Nup107 subcomplex (Radu et al, 1994;Fontoura et al, 1999;Belgareh et al, 2001;, the Nup93-Nup188-Nup205 subcomplex (Grandi et al, 1997;Miller et al, 2000), and a 267-kDa protein termed Tpr (Byrd et al, 1994;Cordes et al, 1997;Zimowska et al, 1997). However, because different antibodies and immunolabeling procedures have been used in different investigations, resulting in partly inconsistent findings, the exact localization of most of these proteins remains unknown.…”

Section: Introductionmentioning

confidence: 99%

Direct Interaction with Nup153 Mediates Binding of Tpr to the Periphery of the Nuclear Pore Complex

Hase

Cordes

2003

MBoC

166

212

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Tpr is a 267-kDa protein forming coiled coil-dominated homodimers that locate at the nucleoplasmic side of the nuclear pore complex (NPC). The proteins that tether Tpr to this location are unknown. Moreover, the question whether Tpr itself might act as a scaffold onto which other NPC components need to be assembled has not been answered to date. To assess Tpr's role as an architectural element of the NPC, we have studied the sequential disassembly and reassembly of NPCs in mitotic cells, paralleled by studies of cells depleted of Tpr as a result of posttranscriptional tpr gene silencing by RNA interference (RNAi). NPC assembly and recruitment of several nucleoporins, including Nup50, Nup93, Nup96, Nup98, Nup107, and Nup153, in anaphase/early telophase is shown to precede NPC association of Tpr in late telophase. In accordance, cellular depletion of Tpr by RNAi does not forestall binding of these nucleoporins to the NPC. In a search for proteins that moor Tpr to the NPC, we have combined the RNAi approach with affinity-chromatography and yeast two-hybrid interaction studies, leading to the identification of nucleoporin Nup153 as the binding partner for Tpr. The specificity of this interaction is demonstrated by its sensitivity to Tpr amino acid substitution mutations that abolish Tpr's ability to adhere to the NPC and affect the direct binding of Tpr to Nup153. Accordingly, cellular depletion of Nup153 by RNAi is shown to result in mislocalization of Tpr to the nuclear interior. Nup153 deficiency also causes mislocalization of Nup50 but has no direct effect on NPC localization of the other nucleoporins studied in this investigation. In summary, these results render Tpr a protein only peripherally attached to the NPC that does not act as an essential scaffold for other nucleoporins

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cDNA Cloning and Characterization ofNpap60:A Novel Rat Nuclear Pore-Associated Protein with an Unusual Subcellular Localization during Male Germ Cell Differentiation

Cited by 40 publications

References 24 publications

The Nucleoporin Nup2 Contains a Meiotic-Autonomous Region that Promotes the Dynamic Chromosome Events of Meiosis

The Nucleoporin Nup2 Contains a Meiotic-Autonomous Region that Promotes the Dynamic Chromosome Events of Meiosis

Characterization and Targeted Disruption of Murine Nup50, a p27^Kip1-Interacting Component of the Nuclear Pore Complex

Direct Interaction with Nup153 Mediates Binding of Tpr to the Periphery of the Nuclear Pore Complex

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cDNA Cloning and Characterization ofNpap60:A Novel Rat Nuclear Pore-Associated Protein with an Unusual Subcellular Localization during Male Germ Cell Differentiation

Cited by 40 publications

References 24 publications

The Nucleoporin Nup2 Contains a Meiotic-Autonomous Region that Promotes the Dynamic Chromosome Events of Meiosis

The Nucleoporin Nup2 Contains a Meiotic-Autonomous Region that Promotes the Dynamic Chromosome Events of Meiosis

Characterization and Targeted Disruption of Murine Nup50, a p27Kip1-Interacting Component of the Nuclear Pore Complex

Direct Interaction with Nup153 Mediates Binding of Tpr to the Periphery of the Nuclear Pore Complex

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Characterization and Targeted Disruption of Murine Nup50, a p27^Kip1-Interacting Component of the Nuclear Pore Complex