A dominant-negative form of POM121 binds chromatin and disrupts the two separate modes of nuclear pore assembly

Shaulov, Lihi; Gruber, Rita; Cohen, I. Bernard; Harel, Amnon

doi:10.1242/jcs.086660

Cited by 36 publications

(28 citation statements)

References 68 publications

(145 reference statements)

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“…5,19,20,64,67,69 LacI-CFP-sPom121, a soluble form of Pom121, when immobilized to the LacO array in our study recruited multiple nucleoporins, including the FG Nup, Nup62 (89% of cells), which was previously observed to be recruited in a study where Pom121 was inserted into the mitochondrial membrane of HeLa and 3T3 cells. 68 LacI-tagged sPom121 also efficiently recruited the FG nucleoporin Nup98 (72% of cells) and the central scaffold protein Nup93 (68% of cells).…”

Section: Soluble Pom121 Cannot Act As An Efficient Seed In the Laci/ supporting

confidence: 57%

Analysis of the initiation of nuclear pore assembly by ectopically targeting nucleoporins to chromatin

Schwartz

Travesa

Martell

et al. 2015

Nucleus

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Nuclear pore complexes (NPCs) form the gateway to the nucleus, mediating virtually all nucleocytoplasmic trafficking. Assembly of a nuclear pore complex requires the organization of many soluble sub-complexes into a final massive structure embedded in the nuclear envelope. By use of a LacI/LacO reporter system, we were able to assess nucleoporin (Nup) interactions, show that they occur with a high level of specificity, and identify nucleoporins sufficient for initiation of the complex process of NPC assembly in vivo. Eleven nucleoporins from different sub-complexes were fused to LacI-CFP and transfected separately into a human cell line containing a stably integrated LacO DNA array. The LacI-Nup fusion proteins, which bound to the array, were examined for their ability to recruit endogenous nucleoporins to the intranuclear LacO site. Many could recruit nucleoporins of the same sub-complex and a number could also recruit other sub-complexes. Strikingly, Nup133 and Nup107 of the Nup107/160 subcomplex and Nup153 and Nup50 of the nuclear pore basket recruited a near full complement of nucleoporins to the LacO array. Furthermore, Nup133 and Nup153 efficiently targeted the LacO array to the nuclear periphery. Our data support a hierarchical, seeded assembly pathway and identify Nup133 and Nup153 as effective "seeds" for NPC assembly. In addition, we show that this system can be applied to functional studies of individual nucleoporin domains as well as to specific nucleoporin disease mutations. We find that the R391H cardiac arrhythmia/sudden death mutation of Nup155 prevents both its subcomplex assembly and nuclear rim targeting of the LacO array.

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Section: Soluble Pom121 Cannot Act As An Efficient Seed In the Laci/ supporting

confidence: 57%

Analysis of the initiation of nuclear pore assembly by ectopically targeting nucleoporins to chromatin

Schwartz

Travesa

Martell

et al. 2015

Nucleus

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“…However, nuclear envelope precursor vesicles crucial to in vitro nuclear assembly, including POM121-containing vesicles, have been found to bind to DNA in the absence of MEL28 (Ulbert et al, 2006). Furthermore, a soluble fragment of POM121 can competitively block nuclear assembly in vitro without disrupting the recruitment of MEL28, and in turn the Nup107-Nup160 complex, owing to distinct binding sites on chromatin (Shaulov et al, 2011). Finally, nuclei assembled in the absence of MEL28 form pore-free, albeit closed, nuclear envelopes (Franz et al, 2007), which we did not observe upon LSD1 depletion.…”

Section: Discussionmentioning

confidence: 53%

The lysine demethylase LSD1 is required for nuclear envelope formation at the end of mitosis

Schooley

Moreno-Andrés

Magistris

et al. 2015

Journal of Cell Science

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The metazoan nucleus breaks down and reassembles during each cell division. Upon mitotic exit, the successful reestablishment of an interphase nucleus requires the coordinated reorganization of chromatin and formation of a functional nuclear envelope. Here, we report that the histone demethylase LSD1 (also known as KDM1A) plays a crucial role in nuclear assembly at the end of mitosis. Downregulation of LSD1 in cells extends telophase and impairs nuclear pore complex assembly. In vitro, LSD1 demethylase activity is required for the recruitment of MEL28 (also known as ELYS and AHCTF1) and nuclear envelope precursor vesicles to chromatin, crucial steps in nuclear reassembly. Accordingly, the formation of a closed nuclear envelope and nuclear pore complex assembly are impaired upon depletion of LSD1 or inhibition of its activity. Our results identify histone demethylation by LSD1 as a new regulatory mechanism linking the chromatin state and nuclear envelope formation at the end of mitosis.

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“…Previous studies suggest that the NLS domain of Pom121 is crucial for interactions with various Nups at the NPC, including Nup98 and the Nup107/ 160 complex (Mitchell et al 2010;Yavuz et al 2010;Shaulov et al 2011). In addition, the NLS domain is required for most of Pom121's described roles at the NPC (Yavuz et al 2010;Shaulov et al 2011). We therefore hypothesized that, like canonical Pom121 at the NPC, sPom121 uses its NLS domain to interact with Nup98 and the Nup107/160 complex except it does so in the nucleoplasm.…”

Section: The Nuclear Localization Signal (Nls) Domain Of Spom121 Is Rmentioning

confidence: 99%

Evolution of a transcriptional regulator from a transmembrane nucleoporin

et al. 2016

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Nuclear pore complexes (NPCs) emerged as nuclear transport channels in eukaryotic cells ∼1.5 billion years ago. While the primary role of NPCs is to regulate nucleo-cytoplasmic transport, recent research suggests that certain NPC proteins have additionally acquired the role of affecting gene expression at the nuclear periphery and in the nucleoplasm in metazoans. Here we identify a widely expressed variant of the transmembrane nucleoporin (Nup) Pom121 (named sPom121, for "soluble Pom121") that arose by genomic rearrangement before the divergence of hominoids. sPom121 lacks the nuclear membrane-anchoring domain and thus does not localize to the NPC. Instead, sPom121 colocalizes and interacts with nucleoplasmic Nup98, a previously identified transcriptional regulator, at gene promoters to control transcription of its target genes in human cells. Interestingly, sPom121 transcripts appear independently in several mammalian species, suggesting convergent innovation of Nup-mediated transcription regulation during mammalian evolution. Our findings implicate alternate transcription initiation as a mechanism to increase the functional diversity of NPC components.

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A dominant-negative form of POM121 binds chromatin and disrupts the two separate modes of nuclear pore assembly

Cited by 36 publications

References 68 publications

Analysis of the initiation of nuclear pore assembly by ectopically targeting nucleoporins to chromatin

Analysis of the initiation of nuclear pore assembly by ectopically targeting nucleoporins to chromatin

The lysine demethylase LSD1 is required for nuclear envelope formation at the end of mitosis

Evolution of a transcriptional regulator from a transmembrane nucleoporin

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