Loss of HCF-1–Chromatin Association Precedes Temperature-Induced Growth Arrest of tsBN67 Cells

Wysocka, Joanna; Reilly, Patrick T.; Herr, Winship

doi:10.1128/mcb.21.11.3820-3829.2001

Cited by 179 publications

(210 citation statements)

References 32 publications

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“…Conversely, on the same nitrocellulose membrane, lamin B, a nuclear matrix protein considered the cornerstone of the insoluble nuclear shell (Vlcek et al, 2001), was predominantly found (ϳ98% of the total amount proceeding from the DNase I-sensitive and nuclear matrix fractions, as measured by densitometry) in the nuclear matrix fraction and barely detected in the DNase I-sensitive fraction (Figure 2A). To confirm that NuMA was present in the chromatin fraction, nuclei were isolated from cells cultured under 2D conditions and treated to separate the chromatin from the nondigestible nuclear remnant according to classical chromatin fractionation methods (Wysocka et al, 2001). Western blot analysis indicated that NuMA was present in the nondigestible nuclear compartment obtained upon micrococcal nuclease treatment, in agreement with previous observations (Lydersen and Pettijohn, 1980;Zeng et al, 1994b).…”

Section: A Fraction Of Numa Is Present In the Chromatin Compartment Dsupporting

confidence: 64%

“…Suspensions were centrifuged at 3200 ϫ g for 15 min at 4°C. The procedure for further isolation of chromatin fractions was performed as described previously (Wysocka et al, 2001) with only minor modifications. Briefly, nuclear pellets were resuspended in buffer X [10 mM HEPES, pH 7.9, 10 mM KCl, 1.5 mM MgCl 2, 0.34 M sucrose, 10% (vol/vol) glycerol, 1 mM dithiothreitol, and PI].…”

Section: Preparation Of Chromatin Fractionsmentioning

confidence: 99%

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NuMA Influences Higher Order Chromatin Organization in Human Mammary Epithelium

Abad

Lewis

Mian

et al. 2007

MBoC

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The coiled-coil protein NuMA is an important contributor to mitotic spindle formation and stabilization. A potential role for NuMA in nuclear organization or gene regulation is suggested by the observations that its pattern of nuclear distribution depends upon cell phenotype and that it interacts and/or colocalizes with transcription factors. To date, the precise contribution of NuMA to nuclear function remains unclear. Previously, we observed that antibody-induced alteration of NuMA distribution in growth-arrested and differentiated mammary epithelial structures (acini) in threedimensional culture triggers the loss of acinar differentiation. Here, we show that in mammary epithelial cells, NuMA is present in both the nuclear matrix and chromatin compartments. Expression of a portion of the C terminus of NuMA that shares sequence similarity with the chromatin regulator HPC2 is sufficient to inhibit acinar differentiation and results in the redistribution of NuMA, chromatin markers acetyl-H4 and H4K20m, and regions of deoxyribonuclease I-sensitive chromatin compared with control cells. Short-term alteration of NuMA distribution with anti-NuMA C-terminus antibodies in live acinar cells indicates that changes in NuMA and chromatin organization precede loss of acinar differentiation. These findings suggest that NuMA has a role in mammary epithelial differentiation by influencing the organization of chromatin. INTRODUCTIONThe nuclear mitotic apparatus protein (NuMA) was first described in 1980 (Lydersen and Pettijohn, 1980) and observed to concentrate at the spindle poles during mitosis. Subsequently NuMA, variously named SPN antigen, p240 antigen, centrophilin, 1F1/1H1 antigen, SP-H antigen, and W1 antigen (Compton et al., 1991(Compton et al., , 1992Kallajoki et al., 1991Kallajoki et al., , 1993Maekawa et al., 1991;Tousson et al., 1991;Yang et al., 1992;Maekawa and Kuriyama, 1993;Tang et al., 1993), was shown to be a critical player in the formation and stabilization of the mitotic spindle, notably by associating with the minus end of spindle microtubules and interacting with dynein, dynactin, and LGN (Compton and Cleveland, 1993;Gaglio et al., 1995;Merdes et al., 2000;Du et al., 2001;Gehmlich et al., 2004). In addition to its location at the poles of the mitotic spindle, NuMA has been reported in the nucleus of both cycling and growth-arrested cells, as shown by immunostaining of cells in culture and tissue biopsy sections (Lelièvre et al., 1998;Merdes and Cleveland 1998;Gribbon et al., 2002;Taimen et al., 2004). However, a role for NuMA in interphase remains to be determined. The hypothesis that NuMA might organize nuclear structure (Compton and Cleveland, 1994) is supported by the existence of a long central coiled-coil region in the protein and the prevalence of NuMA in the nucleus upon detergent extraction. Furthermore, NuMA binds DNA matrix attachment regions (MARs) in vitro (Ludérus et al., 1994), and the cleavage of NuMA precedes DNA degradation during apoptosis (Weaver et al., 1996). The likelihood of a link...

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Section: A Fraction Of Numa Is Present In the Chromatin Compartment Dsupporting

confidence: 64%

Section: Preparation Of Chromatin Fractionsmentioning

confidence: 99%

NuMA Influences Higher Order Chromatin Organization in Human Mammary Epithelium

Abad

Lewis

Mian

et al. 2007

MBoC

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“…Cells were collected after centrifuging at 300 ϫ g for 5 min and washed with cold PBS containing all of the above inhibitors. Cells were fractionated according to previously established methods (28,43) with some modifications. In brief, cells were lysed in 10 mM Hepes buffer (pH 7.6), 10 mM KCl, 1.5 mM MgCl2, 0.34 M sucrose, 10% glycerol, 1 mM DTT, and 0.1% Nonidet P-40 in the presence of all above inhibitors supplemented with 1 mM PMSF and protease inhibitor mixture (Roche) and were incubated on ice for 10 min.…”

Section: Methodsmentioning

confidence: 99%

TGF-β and IL-6 signals modulate chromatin binding and promoter occupancy by acetylated FOXP3

Samanta

Li²,

Song³

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

143

119

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“…2B) without the complication of indirect effects associated with overexpression of SUMO or SUMO-conjugating enzymes. SUMO-H4 fusion constructs were transfected into 293 cells, and the cells were fractionated into soluble (S) and chromatin (C) fractions (26,35). As shown in Fig.…”

Section: Present Inmentioning

confidence: 99%

Histone sumoylation is associated with transcriptional repression

Shiio

Eisenman

2003

Proc. Natl. Acad. Sci. U.S.A.

612

444

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Histone proteins are subject to modifications, such as acetylation, methylation, phosphorylation, ubiquitination, glycosylation, and ADP ribosylation, some of which are known to play important roles in the regulation of chromatin structure and function. Here we report that histone H4 is modified by small ubiquitin-related modifier (SUMO) family proteins both in vivo and in vitro. H4 binds to the SUMO-conjugating enzyme (E2), UBC9, and can be sumoylated in an E1 (SUMO-activating enzyme)-and E2-dependent manner. We present evidence suggesting that histone sumoylation mediates gene silencing through recruitment of histone deacetylase and heterochromatin protein 1.C hromatin is composed of nucleosomes in which 146 bp of DNA are wrapped around a core histone octamer. The octamer consists of a (H3-H4) 2 tetramer associated with two H2A-H2B dimers. The C-terminal globular domains of core histones bind to each other to form the octamer whereas core histone N-terminal tails protrude from the nucleosome and are accessible to the enzymatic modification machineries that catalyze acetylation, methylation, phosphorylation, glycosylation, and ADP ribosylation (for reviews see refs. 1 and 2). Over the last several years, rapid progress has been made in the field of histone modification, owing largely to the discoveries that histone acetyltransferases and histone deacetylases function as transcriptional coactivators and corepressors, respectively (3). These discoveries were followed by analyses of the mechanism and effects of acetylation and the identification of other histone modifications including phosphorylation, methylation, and ubiquitination as regulators of transcription.Small ubiquitin-related modifier (SUMO) is the bestcharacterized member of a growing family of ubiquitin-like proteins involved in posttranslational modifications (for reviews see refs. 4-6). In mammals, there are three members of the SUMO protein family: SUMO-1, SUMO-2 (SMT3a), and SUMO-3 (SMT3b), which are implicated in partly overlapping, yet distinct functions (7,8). SUMO is covalently attached to other proteins through the activities of an enzyme cascade (E1-E2-E3) similar to that for ubiquitination. There is only one known SUMO-activating enzyme, E1 (a heterodimer of SAE1 and SAE2) and only one known SUMO-conjugating enzyme, E2 (UBC9). There appear to be a number of different SUMO ligases (E3s) in higher eukaryotes such as PIAS family proteins (9-11), RanBP2 (12), and the polycomb protein Pc2 (13). To date, dozens of proteins from different species have been identified as sumoylation substrates. Among these are the RanGTPase-activating protein, RanGAP1 (14), PML (15), I B␣ (16), p53 (17, 18), and MDM2 (19). Unlike ubiquitination, sumoylation of proteins has not been linked to protein degradation. Proposed functions for sumoylation include regulation of protein-protein interaction and localization, inhibition of ubiquitin-mediated degradation, and enhancement of transcriptional activity.Histone proteins have been long known to be ubiquitinated (20),...

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Loss of HCF-1–Chromatin Association Precedes Temperature-Induced Growth Arrest of tsBN67 Cells

Cited by 179 publications

References 32 publications

NuMA Influences Higher Order Chromatin Organization in Human Mammary Epithelium

NuMA Influences Higher Order Chromatin Organization in Human Mammary Epithelium

TGF-β and IL-6 signals modulate chromatin binding and promoter occupancy by acetylated FOXP3

Histone sumoylation is associated with transcriptional repression

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