Hanley ML , Yoo TY , Sonnett M , Needleman DJ , Mitchison TJ .

F31 GM116451 NIGMS NIH HHS , R01 GM039565 NIGMS NIH HHS , R37 GM039565 NIGMS NIH HHS

	FIGURE 1:. Phosphatase inhibition activates AURKB and causes a change in the hydrodynamic properties of its associated protein complex. (A) Sequences of the activation loop of human and X. laevis AURKB and X. laevis AURKA. Differences are highlighted in red, and the key phosphorylated threonine is highlighted in turquoise. (B) Mitotic HSS was incubated with kinase inhibitors and okadaic acid. Equal reaction volumes were run on an SDS–PAGE gel and immunoblotted with antibodies against pT232-AURK (which recognizes AURKA, AURKB, and AURKC phosphorylated at the conserved threonine on the activation loop of the kinase), STMN, and α-tubulin. The blot was quantified by normalizing pAURKB intensity to α-tubulin intensity and set such that the lowest value of each replicate was the DMSO control (lanes where no intensity could be detected are marked as zero). The p value between the DMSO and okadaic acid samples is 0.014; n = 3. (C) Mitotic HSS was incubated with kinase inhibitor or DMSO for 25 min, followed by okadaic acid or DMSO for an additional 25 min. Equal volumes were sedimented on 5–40% sucrose gradients for 6 h at 237,000 × g and 4°C. The indicated fractions were separated on SDS–PAGE gels and immunoblotted with antibodies against INCENP and CDCA9. The identity of the higher–molecular weight band in higher-numbered INCENP blots is unknown but likely corresponds to a posttranslationally modified version of INCENP. Full blots with molecular weight markers are given in Supplemental Figure S1. (D) Sucrose gradient blots in C were quantified and normalized to the total amount of the indicated protein in the gradient. Error bars represent SD. The p values indicated by asterisks were calculated between the barasertib- and okadaic acid–treated samples in fractions 3 (INCENP 0.00046, CDCA9 0.0075) and 4 (INCENP 0.0057, CDCA9 0.0062); n = 4.
	FIGURE 2:. CPC hydrodynamic properties are similar in interphase and mitotic HSS. (A) Interphase HSS was prepared by cycling CSF X. laevis egg extract into interphase with calcium and then adding cycloheximide, followed by the high-speed spin. Equal amounts of interphase and mitotic HSS prepared from the same extract were blotted with an MPM2 (mitotic phosphoprotein monoclonal) antibody that recognizes many CDK1 phosphosites more prevalent in mitosis (Kuang et al., 1994; Field et al., 2014). Interphase extract was prepared more than five times and compared with mitotic HSS prepared from the same crude extract using MPM2 blots twice. (B) Sucrose gradients were run as in Figure 1. Mitotic and interphase HSS were prepared from the same extract. Fractions were immunoblotted with antibodies raised against INCENP and CDCA9 after SDS–PAGE. Full blots with molecular weight markers are given in Supplemental Figure S3. (C) Sucrose gradient blots from B were quantified as in Figure 1D. Error bars represent SD; n = 2. For fraction 4, significant p values comparing basal interphase HSS and interphase HSS dosed with 1 and 10 μM okadaic acid (OA) were calculated to be 0.011 (INCENP, 1 μM OA), 0.050 (INCENP, 10 μM OA), and 0.014 (CDCA9, 10 μM OA); n = 2–5. (D) Mitotic and interphase HSS samples prepared from the same extract were incubated with the indicated drugs as in Figure 1. The reactions were then blotted with antibodies against pT232-AURKB and α-tubulin, and pAURKB levels were quantified by normalizing to α-tubulin; n = 2.
	FIGURE 3:. Tagging of CDCA8 in HeLa cells with GFP-3xFLAG. (A) HeLa cells were arrested in mitosis by overnight treatment with STLC and collected via mitotic shake-off, followed by washing with PBS. The indicated drugs were added to the final wash of the cells, after which the cells were lysed with freeze–thaw cycles. Cell lysates were then immunoblotted for pT232-AURK (recognizing pAURKA, pAURKB, and pAURKC) and α-tubulin. The p value between the barasertib- and okadaic acid–treated cells is 0.069; n = 2. (B) Lysates of the parent and tagged cell line were prepared as in A. Magnetic beads preloaded with an antibody against FLAG were used to immunoprecipitate (IP) tagged proteins from the lysates, and the resulting proteins were blotted for FLAG after separation via SDS–PAGE gel electrophoresis; n = 2. (C) Lysates of the tagged cell line were prepared as in A, and IPs were done with beads loaded with antibodies against random IgG or FLAG, followed by blotting for FLAG as in A; n = 2. (D) HeLa cells expressing GFP-tagged CPC were fixed with 4% paraformaldehyde, stained with an antibody against tubulin, and imaged with a spinning-disk confocal microscope at 60× magnification. Scale bars, 10 μm. Cells were imaged on three separate occasions.
	FIGURE 4:. Fluorescence correlation spectroscopy of HeLa CDCA8-GFP-3xFLAG cells. Mitotic cells were identified and CPC localization to centrosomes confirmed by conventional imaging. The FCS measurement beam was directed into the cytoplasm near the edge of cells away from chromosomes to report on soluble CPC diffusion. (A) Example autocorrelation curves (top; black circles represent averages, and red and blue lines represent the fitted one- and two-component FCS models, respectively) and weighted residuals (bottom). (B) Ratio of amplitudes of slow- to fast-diffusing populations over time after drug treatment. Amplitudes are proportional to the number of molecules in a focal volume and molecular brightness squared. (C) Diffusion coefficients over time after drug treatment of the slow (left) and fast (right) populations in the presence of 1 µM nocodazole, with no further drug treatment (blue circles), or after treatment with only 250 nM okadaic acid (OA; green squares) or with both 250 nM OA and 250 nM barasertib (BA; red triangles). Each data point represents one mitotic cell, identified by observing round cell morphology and chromosomes in phase contrast, and averages were obtained by taking multiple measurements in each cell at each time point. Error bars represent the SD of the fitted parameters of the nonlinear regression.
	FIGURE 5:. NPM2 interacts with inactive CPC. (A) CPC proteins were immunoprecipitated from mitotic HSS and analyzed via quantitative mass spectrometry. Protein counts were normalized to IgG IPs and the target protein (INCENP or CDCA9). Ratios for each protein were generated by dividing reporter ion counts from barasertib-treated samples by counts from okadaic acid–treated samples. Proteins with a ratio >1 (gridlines on plot) indicate enrichment in barasertib-treated samples over okadaic acid–treated samples. Labeled proteins are those with ratios >1 in both CDCA8 and INCENP IPs. Size of bubbles is proportional to the measured amount of protein in X. laevis extract (Wuhr et al., 2014). Full lists of proteins and ratios are given in Supplemental Table S1. (B) Mitotic HSS was incubated with barasertib (BA), okadaic acid (OA), both, or neither as in Figure 1. Beads loaded with antibodies against X. laevis INCENP, CDCA9, and NPM2 were used for immunoprecipitations from treated mitotic HSS, and then the proteins on the beads were blotted with antibodies against INCENP and NPM2. Blots were quantified by dividing levels of the blotted protein by levels of the IP target protein. Fold change represents change in NPM2 levels (INCENP and CDCA9 IPs) or INCENP levels (NPM2 IP) after intensities were normalized by setting the DMSO lane to 1. Values of 0 indicate no detectable intensity. The p values between the BA and OA samples indicated by the asterisk are 0.029 (CDCA9 IP) and 0.043 (NPM2 IP); n = 2. Full blots, including molecular weight markers, are given in Supplemental Figure S6. (C) Crude X. laevis extract was cycled into interphase with the addition of calcium, and both mitotic and interphase extract were treated with drugs as in B. Beads loaded with antibody against CDCA9 were used for immunoprecipitation and immunoblotted for INCENP and NPM2. NPM2 blots were quantified as described, normalized to INCENP, and with the lane treated with both drugs set to 1. Full blot including molecular weight markers is given in Supplemental Figure S7. (D) The CPC was immunoprecipitated from mitotic HSS using the antibody raised against CDCA9. After four quick low-salt washes, beads were incubated in solutions of increasing salt for 5 min each. The proteins bound to the beads before and after the salt washes, as well as those in the entire volume of each salt wash, were separated via SDS–PAGE and protein levels analyzed by immunoblots of INCENP, NPM2, and actin. Protein levels were quantified as a percentage of the total of that protein. Error bars represent the SD; n = 2. Full blots including molecular weight markers are given in Supplemental Figure S8. (E) Sucrose gradients of barasertib (BA)- or okadaic acid (OA)–treated mitotic HSS were run as in Figure 1, except that fractions were one-third smaller. Every other fraction was immunoblotted for NPM2, INCENP, or CDCA9. NPM2 blots had 7.5 times less total protein concentration in each lane. NPM2 blots were quantified as in Figure 1. Error bars represent SD; n = 2. Full blots including molecular weight markers are given in Supplemental Figure S9. (F) Proposed model for phosphorylation-induced CPC hydrodynamic changes (CPC proteins: AURKB green, INCENP black line, CDCA8/9 orange, BIRC5 yellow). Pentameric or decameric NPM oligomers (blue) interact with the CPC when the CPC is inactive and unphosphorylated. This interaction may involve direct binding or be mediated by other proteins. When phosphatases are inhibited, NPMs dissociate and the CPC is activated.
	Figure S2: Mitotic HSS was diluted 1, 2, 3, 4, or 5-fold in S-CSF-XB-phosphate buffer, okadaic acid was added at a constant concentration, and the reactions were incubated at room temperature for 25 minutes. Reaction aliquots containing the same amount of HSS were immunoblotted with the antibody against the pT232-AURK epitope. The intensity of the samples was normalized to α-tubulin intensity and across biological replicates, setting the lowest value to one. Error bars represent standard deviations, and the p-value between 100% and 20% HSS is 0.0019, n=4.
	Figure S4. (A) Samples of mitotic or interphase crude X. laevis extract were immunoblotted using an antibody against MPM2 epitopes. (B) Droplets of interphase or mitotic extract (1 μL) from the same extract were placed into a room temperature mineral oil bath and observed via dark field microscopy for a period of approximately 45 minutes.
	Figure S10. Mitotic HSS was treated with no drug, barasertib, okadaic acid, or both, and then Dynabeads conjugated with antibody against CDCA9 were used to immunoprecipitate the CPC. The beads were then immunoblotted for INCENP and histone H2A.Z.
	FIGURE 1:. Phosphatase inhibition activates AURKB and causes a change in the hydrodynamic properties of its associated protein complex. (A) Sequences of the activation loop of human and X. laevis AURKB and X. laevis AURKA. Differences are highlighted in red, and the key phosphorylated threonine is highlighted in turquoise. (B) Mitotic HSS was incubated with kinase inhibitors and okadaic acid. Equal reaction volumes were run on an SDS–PAGE gel and immunoblotted with antibodies against pT232-AURK (which recognizes AURKA, AURKB, and AURKC phosphorylated at the conserved threonine on the activation loop of the kinase), STMN, and α-tubulin. The blot was quantified by normalizing pAURKB intensity to α-tubulin intensity and set such that the lowest value of each replicate was the DMSO control (lanes where no intensity could be detected are marked as zero). The p value between the DMSO and okadaic acid samples is 0.014; n = 3. (C) Mitotic HSS was incubated with kinase inhibitor or DMSO for 25 min, followed by okadaic acid or DMSO for an additional 25 min. Equal volumes were sedimented on 5–40% sucrose gradients for 6 h at 237,000 × g and 4°C. The indicated fractions were separated on SDS–PAGE gels and immunoblotted with antibodies against INCENP and CDCA9. The identity of the higher–molecular weight band in higher-numbered INCENP blots is unknown but likely corresponds to a posttranslationally modified version of INCENP. Full blots with molecular weight markers are given in Supplemental Figure S1. (D) Sucrose gradient blots in C were quantified and normalized to the total amount of the indicated protein in the gradient. Error bars represent SD. The p values indicated by asterisks were calculated between the barasertib- and okadaic acid–treated samples in fractions 3 (INCENP 0.00046, CDCA9 0.0075) and 4 (INCENP 0.0057, CDCA9 0.0062); n = 4.
	FIGURE 2:. CPC hydrodynamic properties are similar in interphase and mitotic HSS. (A) Interphase HSS was prepared by cycling CSF X. laevis egg extract into interphase with calcium and then adding cycloheximide, followed by the high-speed spin. Equal amounts of interphase and mitotic HSS prepared from the same extract were blotted with an MPM2 (mitotic phosphoprotein monoclonal) antibody that recognizes many CDK1 phosphosites more prevalent in mitosis (Kuang et al., 1994; Field et al., 2014). Interphase extract was prepared more than five times and compared with mitotic HSS prepared from the same crude extract using MPM2 blots twice. (B) Sucrose gradients were run as in Figure 1. Mitotic and interphase HSS were prepared from the same extract. Fractions were immunoblotted with antibodies raised against INCENP and CDCA9 after SDS–PAGE. Full blots with molecular weight markers are given in Supplemental Figure S3. (C) Sucrose gradient blots from B were quantified as in Figure 1D. Error bars represent SD; n = 2. For fraction 4, significant p values comparing basal interphase HSS and interphase HSS dosed with 1 and 10 μM okadaic acid (OA) were calculated to be 0.011 (INCENP, 1 μM OA), 0.050 (INCENP, 10 μM OA), and 0.014 (CDCA9, 10 μM OA); n = 2–5. (D) Mitotic and interphase HSS samples prepared from the same extract were incubated with the indicated drugs as in Figure 1. The reactions were then blotted with antibodies against pT232-AURKB and α-tubulin, and pAURKB levels were quantified by normalizing to α-tubulin; n = 2.
	FIGURE 3:. Tagging of CDCA8 in HeLa cells with GFP-3xFLAG. (A) HeLa cells were arrested in mitosis by overnight treatment with STLC and collected via mitotic shake-off, followed by washing with PBS. The indicated drugs were added to the final wash of the cells, after which the cells were lysed with freeze–thaw cycles. Cell lysates were then immunoblotted for pT232-AURK (recognizing pAURKA, pAURKB, and pAURKC) and α-tubulin. The p value between the barasertib- and okadaic acid–treated cells is 0.069; n = 2. (B) Lysates of the parent and tagged cell line were prepared as in A. Magnetic beads preloaded with an antibody against FLAG were used to immunoprecipitate (IP) tagged proteins from the lysates, and the resulting proteins were blotted for FLAG after separation via SDS–PAGE gel electrophoresis; n = 2. (C) Lysates of the tagged cell line were prepared as in A, and IPs were done with beads loaded with antibodies against random IgG or FLAG, followed by blotting for FLAG as in A; n = 2. (D) HeLa cells expressing GFP-tagged CPC were fixed with 4% paraformaldehyde, stained with an antibody against tubulin, and imaged with a spinning-disk confocal microscope at 60× magnification. Scale bars, 10 μm. Cells were imaged on three separate occasions.
	FIGURE 4:. Fluorescence correlation spectroscopy of HeLa CDCA8-GFP-3xFLAG cells. Mitotic cells were identified and CPC localization to centrosomes confirmed by conventional imaging. The FCS measurement beam was directed into the cytoplasm near the edge of cells away from chromosomes to report on soluble CPC diffusion. (A) Example autocorrelation curves (top; black circles represent averages, and red and blue lines represent the fitted one- and two-component FCS models, respectively) and weighted residuals (bottom). (B) Ratio of amplitudes of slow- to fast-diffusing populations over time after drug treatment. Amplitudes are proportional to the number of molecules in a focal volume and molecular brightness squared. (C) Diffusion coefficients over time after drug treatment of the slow (left) and fast (right) populations in the presence of 1 µM nocodazole, with no further drug treatment (blue circles), or after treatment with only 250 nM okadaic acid (OA; green squares) or with both 250 nM OA and 250 nM barasertib (BA; red triangles). Each data point represents one mitotic cell, identified by observing round cell morphology and chromosomes in phase contrast, and averages were obtained by taking multiple measurements in each cell at each time point. Error bars represent the SD of the fitted parameters of the nonlinear regression.
	FIGURE 5:. NPM2 interacts with inactive CPC. (A) CPC proteins were immunoprecipitated from mitotic HSS and analyzed via quantitative mass spectrometry. Protein counts were normalized to IgG IPs and the target protein (INCENP or CDCA9). Ratios for each protein were generated by dividing reporter ion counts from barasertib-treated samples by counts from okadaic acid–treated samples. Proteins with a ratio >1 (gridlines on plot) indicate enrichment in barasertib-treated samples over okadaic acid–treated samples. Labeled proteins are those with ratios >1 in both CDCA8 and INCENP IPs. Size of bubbles is proportional to the measured amount of protein in X. laevis extract (Wuhr et al., 2014). Full lists of proteins and ratios are given in Supplemental Table S1. (B) Mitotic HSS was incubated with barasertib (BA), okadaic acid (OA), both, or neither as in Figure 1. Beads loaded with antibodies against X. laevis INCENP, CDCA9, and NPM2 were used for immunoprecipitations from treated mitotic HSS, and then the proteins on the beads were blotted with antibodies against INCENP and NPM2. Blots were quantified by dividing levels of the blotted protein by levels of the IP target protein. Fold change represents change in NPM2 levels (INCENP and CDCA9 IPs) or INCENP levels (NPM2 IP) after intensities were normalized by setting the DMSO lane to 1. Values of 0 indicate no detectable intensity. The p values between the BA and OA samples indicated by the asterisk are 0.029 (CDCA9 IP) and 0.043 (NPM2 IP); n = 2. Full blots, including molecular weight markers, are given in Supplemental Figure S6. (C) Crude X. laevis extract was cycled into interphase with the addition of calcium, and both mitotic and interphase extract were treated with drugs as in B. Beads loaded with antibody against CDCA9 were used for immunoprecipitation and immunoblotted for INCENP and NPM2. NPM2 blots were quantified as described, normalized to INCENP, and with the lane treated with both drugs set to 1. Full blot including molecular weight markers is given in Supplemental Figure S7. (D) The CPC was immunoprecipitated from mitotic HSS using the antibody raised against CDCA9. After four quick low-salt washes, beads were incubated in solutions of increasing salt for 5 min each. The proteins bound to the beads before and after the salt washes, as well as those in the entire volume of each salt wash, were separated via SDS–PAGE and protein levels analyzed by immunoblots of INCENP, NPM2, and actin. Protein levels were quantified as a percentage of the total of that protein. Error bars represent the SD; n = 2. Full blots including molecular weight markers are given in Supplemental Figure S8. (E) Sucrose gradients of barasertib (BA)- or okadaic acid (OA)–treated mitotic HSS were run as in Figure 1, except that fractions were one-third smaller. Every other fraction was immunoblotted for NPM2, INCENP, or CDCA9. NPM2 blots had 7.5 times less total protein concentration in each lane. NPM2 blots were quantified as in Figure 1. Error bars represent SD; n = 2. Full blots including molecular weight markers are given in Supplemental Figure S9. (F) Proposed model for phosphorylation-induced CPC hydrodynamic changes (CPC proteins: AURKB green, INCENP black line, CDCA8/9 orange, BIRC5 yellow). Pentameric or decameric NPM oligomers (blue) interact with the CPC when the CPC is inactive and unphosphorylated. This interaction may involve direct binding or be mediated by other proteins. When phosphatases are inhibited, NPMs dissociate and the CPC is activated.

Adams, INCENP binds the Aurora-related kinase AIRK2 and is required to target it to chromosomes, the central spindle and cleavage furrow. 2000, Pubmed, Xenbase

Adams, INCENP binds the Aurora-related kinase AIRK2 and is required to target it to chromosomes, the central spindle and cleavage furrow. 2000, Pubmed , Xenbase
Afonso, Feedback control of chromosome separation by a midzone Aurora B gradient. 2014, Pubmed
Arcovito, Synergic role of nucleophosmin three-helix bundle and a flanking unstructured tail in the interaction with G-quadruplex DNA. 2014, Pubmed
Bañuelos, Recognition of intermolecular G-quadruplexes by full length nucleophosmin. Effect of a leukaemia-associated mutation. 2013, Pubmed
Berrabah, Control of nuclear receptor activities in metabolism by post-translational modifications. 2011, Pubmed
Bishop, Phosphorylation of the carboxyl terminus of inner centromere protein (INCENP) by the Aurora B Kinase stimulates Aurora B kinase activity. 2002, Pubmed
Bolli, Born to be exported: COOH-terminal nuclear export signals of different strength ensure cytoplasmic accumulation of nucleophosmin leukemic mutants. 2007, Pubmed
Bolton, Aurora B kinase exists in a complex with survivin and INCENP and its kinase activity is stimulated by survivin binding and phosphorylation. 2002, Pubmed , Xenbase
Bouleau, Maternally inherited npm2 mRNA is crucial for egg developmental competence in zebrafish. 2014, Pubmed
Burns, Roles of NPM2 in chromatin and nucleolar organization in oocytes and embryos. 2003, Pubmed , Xenbase
Carmena, The chromosomal passenger complex (CPC): from easy rider to the godfather of mitosis. 2012, Pubmed
Chiarella, Nucleophosmin mutations alter its nucleolar localization by impairing G-quadruplex binding at ribosomal DNA. 2013, Pubmed
Cong, Multiplex genome engineering using CRISPR/Cas systems. 2013, Pubmed
Cooke, The inner centromere protein (INCENP) antigens: movement from inner centromere to midbody during mitosis. 1987, Pubmed
Cutts, Defective chromosome segregation, microtubule bundling and nuclear bridging in inner centromere protein gene (Incenp)-disrupted mice. 1999, Pubmed
D, Src: more than the sum of its parts. 1997, Pubmed
Dhanasekaran, Multifunctional human transcriptional coactivator protein PC4 is a substrate of Aurora kinases and activates the Aurora enzymes. 2016, Pubmed
Di Natale, Nucleophosmin contains amyloidogenic regions that are able to form toxic aggregates under physiological conditions. 2015, Pubmed
Duan-Porter, Dynamic conformations of nucleophosmin (NPM1) at a key monomer-monomer interface affect oligomer stability and interactions with granzyme B. 2014, Pubmed
Federici, Nucleophosmin C-terminal leukemia-associated domain interacts with G-rich quadruplex forming DNA. 2010, Pubmed
Fernández-Rivero, A Quantitative Characterization of Nucleoplasmin/Histone Complexes Reveals Chaperone Versatility. 2016, Pubmed , Xenbase
Field, Xenopus egg cytoplasm with intact actin. 2014, Pubmed , Xenbase
Field, Actin behavior in bulk cytoplasm is cell cycle regulated in early vertebrate embryos. 2011, Pubmed , Xenbase
Field, Spindle-to-cortex communication in cleaving, polyspermic Xenopus eggs. 2015, Pubmed , Xenbase
Finn, Vertebrate nucleoplasmin and NASP: egg histone storage proteins with multiple chaperone activities. 2012, Pubmed
Gadad, The multifunctional protein nucleophosmin (NPM1) is a human linker histone H1 chaperone. 2011, Pubmed
Gadea, Aurora B is required for mitotic chromatin-induced phosphorylation of Op18/Stathmin. 2006, Pubmed , Xenbase
Gallo, Structure of nucleophosmin DNA-binding domain and analysis of its complex with a G-quadruplex sequence from the c-MYC promoter. 2012, Pubmed
Gassmann, Borealin: a novel chromosomal passenger required for stability of the bipolar mitotic spindle. 2004, Pubmed
Gibson, Enzymatic assembly of DNA molecules up to several hundred kilobases. 2009, Pubmed
Grisendi, Role of nucleophosmin in embryonic development and tumorigenesis. 2005, Pubmed
Groen, Microtubule assembly in meiotic extract requires glycogen. 2011, Pubmed , Xenbase
Haindl, The nucleolar SUMO-specific protease SENP3 reverses SUMO modification of nucleophosmin and is required for rRNA processing. 2008, Pubmed
Hisaoka, Intrinsically disordered regions of nucleophosmin/B23 regulate its RNA binding activity through their inter- and intra-molecular association. 2014, Pubmed
Honda, Exploring the functional interactions between Aurora B, INCENP, and survivin in mitosis. 2003, Pubmed
Hu, Plk1 negatively regulates PRC1 to prevent premature midzone formation before cytokinesis. 2012, Pubmed
Inoue, Involvement of mouse nucleoplasmin 2 in the decondensation of sperm chromatin after fertilization. 2011, Pubmed
Jeyaprakash, Structure of a Survivin-Borealin-INCENP core complex reveals how chromosomal passengers travel together. 2007, Pubmed
Kelly, Chromosomal enrichment and activation of the aurora B pathway are coupled to spatially regulate spindle assembly. 2007, Pubmed , Xenbase
Kim, Fluorescence correlation spectroscopy in living cells. 2007, Pubmed
Koike, Recruitment of phosphorylated NPM1 to sites of DNA damage through RNF8-dependent ubiquitin conjugates. 2010, Pubmed
Koryakina, Androgen receptor phosphorylation: biological context and functional consequences. 2014, Pubmed
Kuang, cdc25 is one of the MPM-2 antigens involved in the activation of maturation-promoting factor. 1994, Pubmed , Xenbase
Kühn, Protein diffusion in mammalian cell cytoplasm. 2011, Pubmed
Laskey, Nucleosomes are assembled by an acidic protein which binds histones and transfers them to DNA. 1978, Pubmed , Xenbase
Lin, Functional characterization and efficient detection of Nucleophosmin/NPM1 oligomers. 2016, Pubmed
Mackay, A dominant mutant of inner centromere protein (INCENP), a chromosomal protein, disrupts prometaphase congression and cytokinesis. 1998, Pubmed
Maggi, Nucleophosmin serves as a rate-limiting nuclear export chaperone for the Mammalian ribosome. 2008, Pubmed
Mitchison, Self-organization of stabilized microtubules by both spindle and midzone mechanisms in Xenopus egg cytosol. 2013, Pubmed , Xenbase
Mitrea, Nucleophosmin integrates within the nucleolus via multi-modal interactions with proteins displaying R-rich linear motifs and rRNA. 2016, Pubmed
Mitrea, Structural polymorphism in the N-terminal oligomerization domain of NPM1. 2014, Pubmed
Mortlock, Discovery, synthesis, and in vivo activity of a new class of pyrazoloquinazolines as selective inhibitors of aurora B kinase. 2007, Pubmed
Mukherjee, Interactions of the natural product (+)-avrainvillamide with nucleophosmin and exportin-1 Mediate the cellular localization of nucleophosmin and its AML-associated mutants. 2015, Pubmed
Namboodiri, The structure and function of Xenopus NO38-core, a histone chaperone in the nucleolus. 2004, Pubmed , Xenbase
Needleman, Pin-hole array correlation imaging: highly parallel fluorescence correlation spectroscopy. 2009, Pubmed
Neo, TRIM28 Is an E3 Ligase for ARF-Mediated NPM1/B23 SUMOylation That Represses Centrosome Amplification. 2015, Pubmed
Nguyen, Spatial organization of cytokinesis signaling reconstituted in a cell-free system. 2014, Pubmed , Xenbase
Okuwaki, Function of homo- and hetero-oligomers of human nucleoplasmin/nucleophosmin family proteins NPM1, NPM2 and NPM3 during sperm chromatin remodeling. 2012, Pubmed
Okuwaki, The structure and functions of NPM1/Nucleophsmin/B23, a multifunctional nucleolar acidic protein. 2008, Pubmed
Okuwaki, Function of nucleophosmin/B23, a nucleolar acidic protein, as a histone chaperone. 2001, Pubmed
Onikubo, Developmentally Regulated Post-translational Modification of Nucleoplasmin Controls Histone Sequestration and Deposition. 2015, Pubmed , Xenbase
Ozlü, Binding partner switching on microtubules and aurora-B in the mitosis to cytokinesis transition. 2010, Pubmed , Xenbase
Petrásek, Precise measurement of diffusion coefficients using scanning fluorescence correlation spectroscopy. 2008, Pubmed
Platonova, Crystal structure and function of human nucleoplasmin (npm2): a histone chaperone in oocytes and embryos. 2011, Pubmed , Xenbase
Portella, Aurora B: a new prognostic marker and therapeutic target in cancer. 2011, Pubmed
Poser, BAC TransgeneOmics: a high-throughput method for exploration of protein function in mammals. 2008, Pubmed
Quensel, In vivo analysis of importin alpha proteins reveals cellular proliferation inhibition and substrate specificity. 2004, Pubmed
Rappsilber, Protocol for micro-purification, enrichment, pre-fractionation and storage of peptides for proteomics using StageTips. 2007, Pubmed
Reboutier, Nucleophosmin/B23 activates Aurora A at the centrosome through phosphorylation of serine 89. 2012, Pubmed
Russo, Insights into amyloid-like aggregation of H2 region of the C-terminal domain of nucleophosmin. 2017, Pubmed
Sagawa, Nucleophosmin deposition during mRNA 3' end processing influences poly(A) tail length. 2011, Pubmed
Samejima, The Inner Centromere Protein (INCENP) Coil Is a Single α-Helix (SAH) Domain That Binds Directly to Microtubules and Is Important for Chromosome Passenger Complex (CPC) Localization and Function in Mitosis. 2015, Pubmed
Sampath, The chromosomal passenger complex is required for chromatin-induced microtubule stabilization and spindle assembly. 2004, Pubmed , Xenbase
Scaloni, Folding mechanism of the C-terminal domain of nucleophosmin: residual structure in the denatured state and its pathophysiological significance. 2009, Pubmed
Scaloni, Deciphering the folding transition state structure and denatured state properties of nucleophosmin C-terminal domain. 2010, Pubmed
Scognamiglio, G-quadruplex DNA recognition by nucleophosmin: new insights from protein dissection. 2014, Pubmed
Sessa, Structure of Aurora B-INCENP in complex with barasertib reveals a potential transinhibitory mechanism. 2014, Pubmed , Xenbase
Sessa, Mechanism of Aurora B activation by INCENP and inhibition by hesperadin. 2005, Pubmed , Xenbase
Shandilya, Phosphorylation of multifunctional nucleolar protein nucleophosmin (NPM1) by aurora kinase B is critical for mitotic progression. 2014, Pubmed
So, G protein-coupled receptor kinase 5 phosphorylates nucleophosmin and regulates cell sensitivity to polo-like kinase 1 inhibition. 2012, Pubmed
Staus, Enhancement of mDia2 activity by Rho-kinase-dependent phosphorylation of the diaphanous autoregulatory domain. 2011, Pubmed
Szebeni, Nucleolar protein B23 has molecular chaperone activities. 1999, Pubmed
Tseng, Dual detection of chromosomes and microtubules by the chromosomal passenger complex drives spindle assembly. 2010, Pubmed , Xenbase
Wilczek, Protein arginine methyltransferase Prmt5-Mep50 methylates histones H2A and H4 and the histone chaperone nucleoplasmin in Xenopus laevis eggs. 2011, Pubmed , Xenbase
Wühr, Deep proteomics of the Xenopus laevis egg using an mRNA-derived reference database. 2014, Pubmed , Xenbase
Wühr, The Nuclear Proteome of a Vertebrate. 2015, Pubmed , Xenbase
Yasui, Autophosphorylation of a newly identified site of Aurora-B is indispensable for cytokinesis. 2004, Pubmed