Centromere Protein (CENP)-W Interacts with Heterogeneous Nuclear Ribonucleoprotein (hnRNP) U and May Contribute to Kinetochore-Microtubule Attachment in Mitotic Cells

Chun, Younghwa; Kim, Raehyung; Lee, Soo Jin

doi:10.1371/journal.pone.0149127

Cited by 16 publications

(12 citation statements)

References 30 publications

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“…anti-H3K4me2 rabbit polyclonal 1/500 Millipore (07-030) [13][14][15] anti-H3K4me3 rabbit polyclonal 1/500 Abcam (ab8580) [16][17][18] anti-H3K9me3 rabbit polyclonal 1/500 Abcam (ab8898) [19][20][21] anti-H3K27ac rabbit polyclonal 1/500 Abcam (ab4729) [22][23][24] anti-H3K27me2 rabbit polyclonal 1/500 Abcam (ab24684) [25,26] anti-H3K27me3 rabbit polyclonal 1/500 Millipore (07-449) [24,[27][28][29] anti-H3K79me2 rabbit polyclonal 1/500 Abcam (ab3594) [30][31][32] anti-H3K79me3 rabbit polyclonal 1/500 Abcam (ab2621) [33][34][35] anti-SCP3 mouse monoclonal 1/500 Abcam (ab97672) [36][37][38] anti-α-Tubulin (S2 Fig) mouse monoclonal 1/500 Sigma-Aldrich (T6199) [39,40] All antibodies are commercially available, and their specificity has been reported in previous studies.…”

Section: Antibody Name Source Dilution Company (Catalogue) Referencementioning

confidence: 99%

Comprehensive histochemical profiles of histone modification in male germline cells during meiosis and spermiogenesis: Comparison of young and aged testes in mice

et al. 2020

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Human epidemiological studies have shown that paternal aging as one of the risk factors for neurodevelopmental disorders, such as autism, in offspring. A recent study has suggested that factors other than de novo mutations due to aging can influence the biology of offspring. Here, we focused on epigenetic alterations in sperm that can influence developmental programs in offspring. In this study, we qualitatively and semiquantitatively evaluated histone modification patterns in male germline cells throughout spermatogenesis based on immunostaining of testes taken from young (3 months old) and aged (12 months old) mice. Although localization patterns were not obviously changed between young and aged testes, some histone modification showed differences in their intensity. Among histone modifications that repress gene expression, histone H3 lysine 9 trimethylation (H3K9me3) was decreased in the male germline cells of the aged testis, while H3K27me2/3 was increased. The intensity of H3K27 acetylation (ac), an active mark, was lower/higher depending on the stages in the aged testis. Interestingly, H3K27ac was detected on the putative sex chromosomes of round spermatids, while other chromosomes were occupied by a repressive mark, H3K27me3. Among other histone modifications that activate gene expression, H3K4me2 was drastically decreased in the male germline cells of the aged testis. In contrast, H3K79me3 was increased in M-phase spermatocytes, where it accumulates on the sex chromosomes. Therefore, aging induced alterations in the amount of histone modifications and in the differences of patterns for each modification. Moreover, histone modifications on the sex chromosomes and on other chromosomes seems to be differentially regulated by aging. These findings will help elucidate the epigenetic mechanisms underlying the influence of paternal aging on offspring development.

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Section: Antibody Name Source Dilution Company (Catalogue) Referencementioning

confidence: 99%

Comprehensive histochemical profiles of histone modification in male germline cells during meiosis and spermiogenesis: Comparison of young and aged testes in mice

et al. 2020

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“…Compared to CENP‐W, which is a small protein consisting of 88 amino acids, hnRNP U is a relatively large protein comprising 806 amino acids. While the N terminus region of hnRNP U is critical for β‐TrCP1 binding , our previous results revealed that its C‐terminal region containing the RGG motif was sufficient for CENP‐W binding . We therefore hypothesized that these three proteins may form a trimeric complex, as depicted in Fig.…”

Section: Resultsmentioning

confidence: 95%

“…Previous studies showed that CENP‐W stably interacts with hnRNP U throughout the cell cycle and that hnRNP U associates with β‐TrCP1 as a pseudosubstrate of the SCF β‐TrCP1 complex . We therefore tested whether CENP‐W also interact with β‐TrCP1.…”

Section: Resultsmentioning

confidence: 99%

“…Centromere protein W (CENP-W) was originally identified as a gene that is commonly activated in human cancer tissues [12] and plays a crucial role in the formation of the kinetochore complex [13,14]. Our recent study revealed that CENP-W interacts with hnRNP U during interphase and mitosis, and mutually increased each other's protein stability by inhibiting proteasome-mediated degradation [15]. In this study, we identified CENP-W as another key regulator of the subcellular localization of b-TrCP1, which may cooperate with hnRNP U.…”

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confidence: 99%

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Centromere protein W interacts with beta‐transducin repeat‐containing protein 1 and modulates its subcellular localization

2016

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Beta-transducin repeat-containing protein 1 (b-TrCP1) is a substrate-recognition module of SCF b-TrCP1 ubiquitin ligases and its subcellular distribution is known to be critical for target specificity. Heterogeneous nuclear ribonucleoprotein (hnRNP) U, an abundant nuclear protein, is known to be a unique regulator of b-TrCP1 shuttling between the cytoplasm and the nucleus. In this study, we report that centromere protein W (CENP-W), which is frequently overexpressed in a variety of human cancers, may also contribute to b-TrCP1 shuttling. Although hnRNP U and CENP-W can interact with b-TrCP1 and transport it independently, these proteins do not compete for b-TrCP1 binding, but rather cooperate to form a stable shuttling complex. Intriguingly, we found that overexpression of CENP-W leads to accumulation of b-TrCP1 in the nucleus. Thus, we propose that CENP-W may function as a booster of b-TrCP1 nuclear import to increase the oncogenicity of b-TrCP1.

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“…Other hnRNP proteins were detected in our data, but because they had abundance ratios <2.75, these proteins were not pursued. hnRNP R and hnRNP U were initially identified as part of a large supramolecular heterogeneous ribonucleoprotein particle, but have also been shown to be associated with other smaller ribonucleoprotein complexes with functions in the nucleus and cytoplasm [38][39][40][41] . To determine if hnRNP R and hnRNP U are part of the same protein complex, we performed immunoprecipitation and immunoblotting with antibodies to the two proteins with extract from HEK293T cells.…”

Section: Identification Of Kpna7 Interacting Proteins In Neuronsmentioning

confidence: 99%

An epilepsy-associated mutation in the nuclear import receptor KPNA7 reduces nuclear localization signal binding

Oostdyk

Wang

Zang

et al. 2020

Sci Rep

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KPNA7 is a member of the Importin-α family of nuclear import receptors. KPNA7 forms a complex with importin-β and facilitates the translocation of signal-containing proteins from the cytoplasm to the nucleus. Exome sequencing of siblings with severe neurodevelopmental defects and clinical features of epilepsy identified two amino acid-altering mutations in KPNA7. Here, we show that the E344Q substitution reduces KPNA7 binding to nuclear localization signals, and that this limits KPNA7 nuclear import activity. The P339A substitution, by contrast, has little effect on KPNA7 binding to nuclear localization signals. Given the neuronal phenotype described in the two patients, we used SILAC labeling, affinity enrichment, and mass spectrometry to identify KPNA7-interacting proteins in human induced pluripotent stem cell-derived neurons. We identified heterogeneous nuclear ribonucleoproteins hnRNP R and hnRNP U as KPNA7-interacting proteins. The E344Q substitution reduced binding and KPNA7mediated import of these cargoes. The c.1030G > C allele which generates E344Q is within a predicted CTCF binding site, and we found that it reduces CTCF binding by approximately 40-fold. Our data support a role for altered neuronal expression and activity of KPNA7 in a rare type of pediatric epilepsy.The bi-directional transport of proteins and RNAs between the cytoplasm and nucleus is one of many pathways that contribute to cellular homeostasis. Dysregulation of nuclear transport has been implicated in diseases including cancer and diverse neurodegenerative disorders 1,2 . This includes disruption of pathways responsible for facilitating nuclear import of proteins that contain a nuclear localization signal (NLS). The composition and sequence of NLSs vary between proteins but consist primarily of either one (monopartite) or two (bipartite) clusters of amino acids enriched in the basic residues lysine and arginine 3,4 . Classical NLS import is mediated by Importin α (Imp-α) and Importin β (Imp-β) 5 . Imp-α functions as an adapter and makes direct contact with both an NLS sequence and Imp-β to facilitate the assembly of a heterotrimeric import complex that shuttles from the cytoplasm to the nucleus 6 . Within the nucleus, RanGTP facilitates release of the NLS containing proteins and it also promotes recycling of the import machinery to the cytoplasm, in preparation for a new round of import 7,8 .In humans, the Imp-α family, which is also known as the karyopherin α family, contains seven members (KPNA1-KPNA7) 9 . These receptors have a highly related protein architecture that includes an N-terminal Imp-β binding (IBB) domain and a super-helical core structure formed by the tandem arrangement of ten armadillo (ARM) repeats 10,11 . A subset of the ARMs are used to create the surfaces for NLS binding. ARMs 2-4 provide the major NLS binding groove for both mono-and bipartite NLS sequences, while ARMs 6-8 generate the minor NLS binding groove for binding the smaller, second cluster of basic residues in bipartite NLSs 12 . Despite the overall s...

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Centromere Protein (CENP)-W Interacts with Heterogeneous Nuclear Ribonucleoprotein (hnRNP) U and May Contribute to Kinetochore-Microtubule Attachment in Mitotic Cells

Cited by 16 publications

References 30 publications

Comprehensive histochemical profiles of histone modification in male germline cells during meiosis and spermiogenesis: Comparison of young and aged testes in mice

Comprehensive histochemical profiles of histone modification in male germline cells during meiosis and spermiogenesis: Comparison of young and aged testes in mice

Centromere protein W interacts with beta‐transducin repeat‐containing protein 1 and modulates its subcellular localization

An epilepsy-associated mutation in the nuclear import receptor KPNA7 reduces nuclear localization signal binding

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