Heterogeneous nuclear ribonucleoprotein C1/C2, MeCP1, and SWI/SNF form a chromatin remodeling complex at the beta-globin locus control region.

Locus control regions (LCRs) are regulatory DNA sequences that are situated many kilobases away from their cognate promoters. LCRs protect transgenes from position effect variegation and heterochromatinization and also promote copy-number dependence of the levels of transgene expression. In this work, we describe the biochemical purification of a previously undescribed LCR-associated remodeling complex (LARC) that consists of heterogeneous nuclear ribonucleoprotein C1/C2, nucleosome remodeling SWI/SNF, and nucleosome remodeling and deacetylating (NuRD)/MeCP1 as a single homogeneous complex. LARC binds to the hypersensitive 2 (HS2)-Maf recognition element (MARE) DNA in a sequence-specific manner and remodels nucleosomes. Heterogeneous nuclear ribonucleoprotein C1/C2, previously known as a general RNA binding protein, provides a sequence-specific DNA recognition element for LARC, and the LARC DNA-recognition sequence is essential for the enhancement of transcription by HS2. Independently of the initiation of transcription, LARC becomes associated with beta-like globin promoters.

[1]  B. Emerson,et al.  A SWI/SNF–Related Chromatin Remodeling Complex, E-RC1, Is Required for Tissue-Specific Transcriptional Regulation by EKLF In Vitro , 1998, Cell.

[2]  H. Erdjument-Bromage,et al.  Tissue-specific and developmental stage-specific DNA binding by a mammalian SWI/SNF complex associated with human fetal-to-adult globin gene switching. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[3]  Jeroen Krijgsveld,et al.  GATA‐1 forms distinct activating and repressive complexes in erythroid cells , 2005, The EMBO journal.

[4]  F. Grosveld,et al.  The beta‐globin dominant control region: hypersensitive site 2. , 1990, The EMBO journal.

[5]  J. Strouboulis,et al.  Heterochromatin Effects on the Frequency and Duration of LCR-Mediated Gene Transcription , 1996, Cell.

[6]  Félix Recillas-Targa,et al.  Position-effect protection and enhancer blocking by the chicken β-globin insulator are separable activities , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[7]  R. Aebersold,et al.  Dynamic changes in transcription factor complexes during erythroid differentiation revealed by quantitative proteomics , 2004, Nature Structural &Molecular Biology.

[8]  J. McDowell,et al.  Essential role of NF-E2 in remodeling of chromatin structure and transcriptional activation of the epsilon-globin gene in vivo by 5' hypersensitive site 2 of the beta-globin locus control region , 1996, Molecular and cellular biology.

[9]  M. Groudine,et al.  The Locus Control Region Is Necessary for Gene Expression in the Human β-Globin Locus but Not the Maintenance of an Open Chromatin Structure in Erythroid Cells , 1998, Molecular and Cellular Biology.

[10]  G. Stamatoyannopoulos,et al.  Hypersensitive site 5 of the human beta locus control region functions as a chromatin insulator. , 1994, Blood.

[11]  J. D. Engel,et al.  Human β-Globin Locus Control Region HS5Contains CTCF- and Developmental Stage-Dependent Enhancer-BlockingActivity in ErythroidCells , 2003, Molecular and Cellular Biology.

[12]  W. C. Forrester,et al.  A deletion of the human beta-globin locus activation region causes a major alteration in chromatin structure and replication across the entire beta-globin locus. , 1990, Genes & development.

[13]  T. Ley,et al.  Targeted deletion of 5'HS2 of the murine beta-globin LCR reveals that it is not essential for proper regulation of the beta-globin locus. , 1995, Genes & development.

[14]  J. Sharpe,et al.  A single beta-globin locus control region element (5' hypersensitive site 2) is sufficient for developmental regulation of human globin genes in transgenic mice , 1992, Molecular and cellular biology.

[15]  W Miller,et al.  Locus control regions of mammalian beta-globin gene clusters: combining phylogenetic analyses and experimental results to gain functional insights. , 1997, Gene.

[16]  T. Kimbrough,et al.  Effect of deletion of 5'HS3 or 5'HS2 of the human beta-globin locus control region on the developmental regulation of globin gene expression in beta-globin locus yeast artificial chromosome transgenic mice. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[17]  J. Stamatoyannopoulos,et al.  NF‐E2 and GATA binding motifs are required for the formation of DNase I hypersensitive site 4 of the human beta‐globin locus control region. , 1995, The EMBO journal.

[18]  H. Erdjument-Bromage,et al.  An Ikaros-Containing Chromatin-Remodeling Complex in Adult-Type Erythroid Cells , 2000, Molecular and Cellular Biology.

[19]  S. Orkin,et al.  Purification of the human NF-E2 complex: cDNA cloning of the hematopoietic cell-specific subunit and evidence for an associated partner , 1993, Molecular and cellular biology.

[20]  J. D. Engel,et al.  Hypersensitive Site 2 Specifies a Unique Function within the Human β-Globin Locus Control Region To Stimulate Globin Gene Transcription , 1999, Molecular and Cellular Biology.

[21]  A. Dean,et al.  A Human Globin Enhancer Causes both Discrete and Widespread Alterations in Chromatin Structure , 2003, Molecular and Cellular Biology.

[22]  W. Hörz,et al.  ATP-dependent nucleosome remodeling. , 2002, Annual review of biochemistry.

[23]  M. Groudine,et al.  The β-Globin LCR Is Not Necessary for an Open Chromatin Structure or Developmentally Regulated Transcription of the Native Mouse β-Globin Locus , 1998 .

[24]  Tony Kouzarides,et al.  Histone H3 Lysine 4 Methylation Disrupts Binding of Nucleosome Remodeling and Deacetylase (NuRD) Repressor Complex* , 2002, The Journal of Biological Chemistry.

[25]  Paul Tempst,et al.  Erythroid transcription factor NF-E2 is a haematopoietic-specific basic–leucine zipper protein , 1993, Nature.

[26]  Q. Feng,et al.  The NuRD complex: linking histone modification to nucleosome remodeling. , 2003, Current topics in microbiology and immunology.

[27]  A. West,et al.  Recruitment of histone modifications by USF proteins at a vertebrate barrier element. , 2004, Molecular cell.

[28]  P. Fraser,et al.  Intergenic transcription and developmental remodeling of chromatin subdomains in the human beta-globin locus. , 2000, Molecular cell.

[29]  B. Emerson,et al.  NF-E2 disrupts chromatin structure at human beta-globin locus control region hypersensitive site 2 in vitro , 1996, Molecular and cellular biology.

[30]  N. Proudfoot,et al.  Definition of transcriptional promoters in the human beta globin locus control region. , 2002, Journal of molecular biology.

[31]  M. Gerstein,et al.  GATA-1 binding sites mapped in the β-globin locus by using mammalian chIp-chip analysis , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[32]  A. Nienhuis,et al.  Tandem AP-1-binding sites within the human beta-globin dominant control region function as an inducible enhancer in erythroid cells. , 1990, Genes & development.

[33]  R. Rosli,et al.  Requiem: a novel zinc finger gene essential for apoptosis in myeloid cells. , 1994, The Journal of biological chemistry.

[34]  M. Groudine,et al.  β-globin Gene Switching and DNase I Sensitivity of the Endogenous β-globin Locus in Mice Do Not Require the Locus Control Region , 2000 .

[35]  R. Kiyama,et al.  Enhancer activity of HS2 of the human beta-LCR is modulated by distance from the key nucleosome. , 2001, Nucleic acids research.

[36]  Xiangdong Fang,et al.  Locus control regions. , 2002, Blood.

[37]  D. Tuan,et al.  Transcription of the HS2 enhancer toward a cis-linked gene is independent of the orientation, position, and distance of the enhancer relative to the gene , 1997, Molecular and cellular biology.

[38]  A. West,et al.  Conserved CTCF Insulator Elements Flank the Mouse and Human β-Globin Loci , 2002, Molecular and Cellular Biology.

[39]  J. Bieker,et al.  Role of Erythroid Kruppel-like Factor in Human - to -Globin Gene Switching (*) , 1995, The Journal of Biological Chemistry.

[40]  G. Blobel,et al.  FOG‐1 recruits the NuRD repressor complex to mediate transcriptional repression by GATA‐1 , 2005, The EMBO journal.

[41]  D. Tuan,et al.  Transcription of the hypersensitive site HS2 enhancer in erythroid cells. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[42]  M. Mahajan,et al.  DNA-dependent adenosine triphosphatase (helicaselike transcription factor) activates beta-globin transcription in K562 cells. , 2002, Blood.

[43]  P. Levings,et al.  The human β‐globin locus control region , 2002 .

[44]  G. Stamatoyannopoulos,et al.  Genomic footprinting and sequencing of human beta-globin locus. Tissue specificity and cell line artifact. , 1994, The Journal of biological chemistry.