Developmental changes in transcription factors associated with the nuclear matrix of chicken erythrocytes

Sun, Jian-Min; Chen, Hou Yu; Litchfield, David W.; Davie, James R.

doi:10.1002/(sici)1097-4644(19960915)62:4<454::aid-jcb3>3.0.co;2-l

Cited by 13 publications

(12 citation statements)

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“…Isolated nuclei from chicken immature erythrocytes were extracted by incubation on ice with 0.3 M NaCl for 10 min. The nuclear extract was the supernatant collected after centrifugation (11).…”

Section: Methodsmentioning

confidence: 99%

Differential Distribution of Unmodified and Phosphorylated Histone Deacetylase 2 in Chromatin

Sun

Chen

Davie

2007

Journal of Biological Chemistry

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Histone deacetylase 2 (HDAC2) is one of the histone-modifying enzymes that regulate gene expression by remodeling chromatin structure. Along with HDAC1, HDAC2 is found in the Sin3 and NuRD multiprotein complexes, which are recruited to promoters by DNA-binding proteins. In this study, we show that the majority of HDAC2 in human breast cancer cells is not phosphorylated. However, the minor population of HDAC2, preferentially cross-linked to DNA by cisplatin, is mono-, di-, or triphosphorylated. Furthermore, HDAC2 phosphorylation is required for formation of Sin3 and NuRD complexes and recruitment to promoters by transcription factors including p53, Rb, YY1, NF-B, Sp1, and Sp3. Unmodified HDAC2 requires linker DNA to associate with chromatin but is not cross-linked to DNA by formaldehyde. We provide evidence that unmodified HDAC2 is associated with the coding region of transcribed genes, whereas phosphorylated HDAC2 is primarily recruited to promoters.Histone-modifying enzymes and ATP-dependent chromatin-remodeling complexes affect gene expression by altering the compaction level of chromatin and the accessibility of DNA to binding proteins. Histone hyperacetylation is generally associated with chromatin decondensation and increased transcriptional activity, whereas histone hypoacetylation contributes to chromatin condensation and transcriptional repression (1, 2). Dynamic histone acetylation at transcriptionally active genes, resulting from the opposing activities of histone deacetylases (HDACs) 2 and histone acetyltransferases, leads to a rapid oscillation between condensed and decondensed chromatin states (3, 4). Four classes of HDACs have been identified in mammalian cells (5). HDAC1 and 2 belong to class I and are homologous to yeast RPD3. Both HDACs are core components of multi-protein corepressor complexes like Sin3 and NuRD, in which their activities are modulated through interactions with other proteins while being recruited by transcription factors to specific promoters (6). HDAC1 and 2 are phosphoproteins, and this post-translational modification enhances their enzymatic activity (7-9). In studies with exogenously expressed tagged HDAC1 and 2, HDAC phosphorylation appeared to be a prerequisite to form the corepressor complexes. However, studies characterizing endogenous HDAC corepressor complexes are lacking.The role of phosphorylation in the recruitment of HDAC2 by transcription factors has not yet been investigated. Several transcription factors repress gene expression by recruiting HDAC1/2 corepressor complexes to the promoters that they affect. Further, pending the promoter context, transcription factors recruit HDAC1 and 2 corepressor complexes to mediate dynamic deacetylation of histones and non-histone chromosomal proteins associated with or close to the promoter. We have previously reported that the Sp1 and Sp3 transcription factors are associated with phosphorylated HDAC2 in breast cancer cells. Although most HDAC2 in breast cancer cells was not phosphorylated, it was the phosphorylated form of...

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Section: Methodsmentioning

confidence: 99%

Differential Distribution of Unmodified and Phosphorylated Histone Deacetylase 2 in Chromatin

Sun

Chen

Davie

2007

Journal of Biological Chemistry

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“…The reaction tube was preincubated at 30°C for 10 min before the addition of purified CK2 (from bovine testis as described previously (18,21)). The reaction was incubated for 10 min before termination upon the addition of SDS sample buffer.…”

Section: Methodsmentioning

confidence: 99%

The Transcriptional Repressor Sp3 Is Associated with CK2-phosphorylated Histone Deacetylase 2

Sun

Chen

Moniwa

et al. 2002

Journal of Biological Chemistry

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Sp1 and Sp3 are ubiquitously expressed mammalian transcription factors that function as activators or repressors. Although both transcription factors share a common domain involved in forming multimers, we demonstrate that Sp1 and Sp3 form separate complexes in estrogen-dependent human breast cancer cells. Sp1 and Sp3 complexes associate with histone deacetylases (HDACs) 1 and 2. Although most HDAC2 is not phosphorylated in the breast cancer cells, HDAC2 bound to Sp1 and Sp3 and cross-linked to chromatin in situ is highly enriched in a phosphorylated form that has a reduced mobility in SDS-polyacrylamide gels. We show that protein kinase CK2 is associated with and phosphorylates HDAC2. Alkaline phosphatase treatment of HDAC2 and Sp1 and Sp3 complexes reduced the associated HDAC activity. Protein kinase CK2 is up-regulated in several cancers including breast cancer, and Sp1 and Sp3 have key roles in estrogen-induced proliferation and gene expression in estrogen-dependent breast cancer cells. CK2 phosphorylation of HDAC2 recruited by Sp1 or Sp3 could regulate HDAC activity and alter the balance of histone deacetylase and histone acetyltransferase activities and dynamic chromatin remodeling of estrogenregulated genes.Remodeling of chromatin structure mediated by ATP-driven chromatin-remodeling complexes and histone-modifying enzymes has a crucial role in gene expression. Acetylation of the core histones favors decondensation of the chromatin fiber by preventing interfiber interactions, whereas the unacetylated histone state contributes to chromatin condensation (1, 2). Dynamic histone acetylation catalyzed by histone deacetylases (HDAC) 1 and histone acetyltransferases allows the chromatin fiber to rapidly oscillate from the condensed and decondensed states (3, 4). In mammalian cells three classes of HDACs are identified. Class I HDACs, such as HDAC1 and HDAC2, are homologous to yeast RPD3, whereas class II HDACs are similar to yeast HDA1. Class III HDACs are related to yeast SIR2 (5, 6). HDAC1 and -2 are components of large multisubunit complexes called Sin3 or NuRD, which are recruited by transcriptional factors such as Mad, YY1, and Rb (5, 7-9).Mammalian cells ubiquitously express Sp1 and Sp3. Sp3 has three isoforms, a long (L-Sp3) and two short forms (M1-Sp3, M2-Sp3) that are the products of differential translational initiation (10). Sp3 may act as a repressor or an activator, with the short forms acting only as repressors (10). The protein structure of L-Sp3 is very similar to that of Sp1, except that Sp3 has a repression domain located N-terminal to the zinc finger DNAbinding domain (11). It has been reported that the relative levels of Sp3 forms change with differentiation, with the differentiated Caco-2 cells expressing more long than short forms (12). Further, alterations in the relative levels of Sp1 to Sp3 have been recorded, with Sp3 levels being greater than Sp1 in primary keratinocytes (13).In this study we investigated the association of histone deacetylase with Sp3 in human breast cancer cells. We ...

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“…The nuclear matrix is selective for which transcription factors it binds, and this selectivity varies with cell type (van Wijnen et al, 1993;1996a). It has been postulated that the nuclear matrix has a role in the expression of genes by concentrating a subset of transcription factors at specific nuclear sites (Stein et al, 1991;Merriman et al, 1995).…”

Section: Nuclear Matrix and Processing Of The Genetic Informationmentioning

confidence: 99%

“…Transcription factors associated with the nuclear matrix can change throughout development and differentiation (Stein et al, 1994;Merriman et al, 1995;Bagchi et al, 1995;Sun et al, 1996a). For example, transcription factors associated with the chick erythrocyte nuclear matrix change throughout development (Sun et al, 1996a).…”

Section: Nuclear Matrix and Processing Of The Genetic Informationmentioning

confidence: 99%

“…For example, transcription factors associated with the chick erythrocyte nuclear matrix change throughout development (Sun et al, 1996a). Primitive red blood cells from 5-day old embryos have high levels of nuclear matrix-bound transcription factors, including GATA-1, CACCC-binding proteins, and NF1; factors that have key roles in erythroid-specific gene expression.…”

Section: Nuclear Matrix and Processing Of The Genetic Informationmentioning

confidence: 99%

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Role of Nuclear Matrix in Estrogen Regulated Gene Expression in Human Breast Cancer Cells

Holth¹

1999

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Public reporting burden for this collection of information is estimated to average 1 hour per response, including the time for reviewing instructions, searching existing data sources, gathering and maintaining the data needed, and completing and reviewing the collection of information. Send comments regarding this burden estimate or any other aspect of this collection of information, including suggestions for reducing this burden, to Washington Headquarters Services, Directorate tor Information Operations and Reports, 1215 Jefferson Davis Highway, Suite 1204, Arlington, VA 22202-4302, and to the Office of Management and Budget, Paperwork Reduction Project (0704-0188), Washington, DC 20503. AGENCY USE ONLY (Leave blank)2. REPORT DATE August 1999 REPORT TYPE AND DATES COVEREDFinal (15 Jul 96 -14 Jul 99) TITLE AND SUBTITLE Role of Nuclear Matrix in Estrogen Regulated Gene Expression in Human Breast Cancer Cells AUTHOR(S)Laurel T, Holth, Ph.D. FUNDING NUMBERS DAMD17-96-1-6269 PERFORMING ORGANIZATION NAME(S) AND ADDRESS(ES)University of Manitoba Winnipeg, Manitoba, Canada R3E-OW3 PERFORMING ORGANIZATION REPORT NUMBER SPONSORING/MONITORING AGENCY NAME(S) AND ADDRESS(ES)Commander ABSTRACT (Maximum 200The goal of this research is to study association of the estrogen receptor a (ER) with the nuclear matrix, and to identify nuclear matrix sites for ER. We constructed an ER fusion protein (GFP-ER) with a His6 tag, HA tag and the green fluorescent protein. Using fluorescent imaging techniques we studied the cellular localization of GFP-ER in the presence of various ER ligands in human breast cancer cell lines transiently transfected to express the GFP-ER protein. Striking differences in the patterns of distribution were observed. Isolation of GFP-ER associated proteins requires a large number of cell expressing GFP-ER. Therefore, a MCF7 stable cell line expressing GFP-ER under the control of an inducible promoter was constructed. Levels of GFP-ER expression can be controlled in this cell line, and this cell line has been carefully analyzed to determine the effect of GFP-ER expression on cell growth and gene expression. We demonstrated that GFP-ER association with the nuclear matrix increases with the length of exposure to estrogen, and that ER associated to the nuclear matrix is also binding to DNA, suggesting that nuclear matrix bound ER is transcriptionally active. SUBJECT TERMS Breast Cancer.Estrogen Receptor, Nuclear Matrix Where copyrighted material is quoted, permission has been obtained to use such material.Where material from documents designated for limited distribution is quoted, permission has been obtained to use the material.Citations of commercial organizations and trade names in this report do not constitute an official Department of Army endorsement or approval of the products or services of these organizations.yln In conducting research using animals, the investigator(s) adhered to the "Guide for the Care and Use of Laboratory Animals," prepared by the Committee on Care and use of Laboratory Animals of the...

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Developmental changes in transcription factors associated with the nuclear matrix of chicken erythrocytes

Cited by 13 publications

References 53 publications

Differential Distribution of Unmodified and Phosphorylated Histone Deacetylase 2 in Chromatin

Differential Distribution of Unmodified and Phosphorylated Histone Deacetylase 2 in Chromatin

The Transcriptional Repressor Sp3 Is Associated with CK2-phosphorylated Histone Deacetylase 2

Role of Nuclear Matrix in Estrogen Regulated Gene Expression in Human Breast Cancer Cells

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