An Sp1/KLF binding site is important for the activity of a Polycomb group response element from the Drosophila engrailed gene

Polycomb-group response elements (PREs) are DNA elements through which the Polycomb-group (PcG) of transcriptional repressors act. Many of the PcG proteins are associated with two protein complexes that repress gene expression by modifying chromatin. Both of these protein complexes specifically associate with PREs in vivo, however, it is not known how they are recruited or held at the PRE. PREs are complex elements, made up of binding sites for many proteins. Our laboratory has been working to define all the sequences and DNA binding proteins required for the activity of a 181 bp PRE from the Drosophila engrailed gene. Here we show that one of the sites necessary for PRE activity, Site 2, can be bound by members of the Sp1/KLF family of zinc finger proteins. There are 10 Sp1/KLF family members in Drosophila, and nine of them bind to Site 2. We derive a consensus binding site for the Sp1/KLF Drosophila family members and show that this consensus sequence is present in most of the molecularly characterized PREs. These data suggest that one or more Sp1/KLF family members play a role in PRE function in Drosophila.

[1]  Mark Rebeiz,et al.  GenePalette: a universal software tool for genome sequence visualization and analysis. , 2004, Developmental biology.

[2]  J. Kassis.  Pairing-sensitive silencing, polycomb group response elements, and transposon homing in Drosophila. , 2002, Advances in genetics.

[3]  R. Tjian,et al.  Transcription factors that activate the Ultrabithorax promoter in developmentally staged extracts , 1988, Cell.

[4]  Thomas Shenk,et al.  Interaction between transcription factors Spl and YY1 , 1993, Nature.

[5]  M. Biggin,et al.  Zeste maintains repression of Ubx transgenes: support for a new model of Polycomb repression. , 2002, Development.

[6]  G. Korge,et al.  The Pipsqueak Protein of Drosophila melanogasterBinds to GAGA Sequences through a Novel DNA-binding Domain* , 1998, The Journal of Biological Chemistry.

[7]  T. Kaufman,et al.  Regulatory regions of the homeotic gene proboscipedia are sensitive to chromosomal pairing. , 1995, Genetics.

[8]  J. Kassis.  Unusual properties of regulatory DNA from the Drosophila engrailed gene: three "pairing-sensitive" sites within a 1.6-kb region. , 1994, Genetics.

[9]  Chih-Chao Yang,et al.  pipsqueak Encodes a Factor Essential for Sequence-Specific Targeting of a Polycomb Group Protein Complex , 2002, Molecular and Cellular Biology.

[10]  Lin Li,et al.  Gene regulation by Sp1 and Sp3. , 2004, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[11]  R. Lehmann,et al.  Identification of genes controlling germ cell migration and embryonic gonad formation in Drosophila. , 1998, Development.

[12]  Stuart S Levine,et al.  Division of labor in polycomb group repression. , 2004, Trends in biochemical sciences.

[13]  Y. Jan,et al.  A Gain-of-Function Screen for Genes That Affect the Development of the Drosophila Adult External Sensory Organ. , 2001, Genetics.

[14]  M. Fujioka,et al.  A complex array of DNA-binding proteins required for pairing-sensitive silencing by a polycomb group response element from the Drosophila engrailed gene. , 2002, Genetics.

[15]  Renato Paro,et al.  Genome-wide prediction of Polycomb/Trithorax response elements in Drosophila melanogaster. , 2003, Developmental cell.

[16]  Brigitte Wild,et al.  Histone Methyltransferase Activity of a Drosophila Polycomb Group Repressor Complex , 2002, Cell.

[17]  H. Jäckle,et al.  A Drosophila homologue of human Sp1 is a head-specific segmentation gene , 1993, Nature.

[18]  P. Schedl,et al.  A Polycomb and GAGA dependent silencer adjoins the Fab-7 boundary in the Drosophila bithorax complex. , 1997, Genetics.

[19]  E. Jane,et al.  The Drosophila Polycomb Group proteins ESC and E(Z) are present in a complex containing the histone-binding protein p55 and the histone deacetylase RPD3. , 2001, Development.

[20]  M. Lehmann,et al.  Pipsqueak and GAGA factor act in concert as partners at homeotic and many other loci , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[21]  M. Whiteley,et al.  The Drosophila Polycomb group gene pleiohomeotic encodes a DNA binding protein with homology to the transcription factor YY1. , 1998, Molecular cell.

[22]  C. Verrijzer,et al.  Characterization of the Extended Myb-like DNA-binding Domain of Trithorax Group Protein Zeste* , 2002, The Journal of Biological Chemistry.

[23]  Jürg Müller,et al.  Molecular and genetic analysis of the Polycomb group gene Sex combs extra/Ring in Drosophila , 2003, Mechanisms of Development.

[24]  F. Karch,et al.  The Fab-8 boundary defines the distal limit of the bithorax complex iab-7 domain and insulates iab-7 from initiation elements and a PRE in the adjacent iab-8 domain. , 2000, Development.

[25]  A. Kullyev,et al.  The Mcp Element from the bithorax Complex Contains an Insulator That Is Capable of Pairwise Interactions and Can Facilitate Enhancer-Promoter Communication , 2005, Molecular and Cellular Biology.

[26]  A. Lund,et al.  Polycomb complexes and silencing mechanisms. , 2004, Current opinion in cell biology.

[27]  R. Paro,et al.  Co‐localization of Polycomb protein and GAGA factor on regulatory elements responsible for the maintenance of homeotic gene expression , 1997, The EMBO journal.

[28]  R. Paro,et al.  Binding of Trithorax and Polycomb proteins to the bithorax complex: dynamic changes during early Drosophila embryogenesis , 1998, The EMBO journal.

[29]  F. Karch,et al.  The iab-7 Polycomb Response Element Maps to a Nucleosome-Free Region of Chromatin and Requires Both GAGA and Pleiohomeotic for Silencing Activity , 2001, Molecular and Cellular Biology.

[30]  C. Fritsch,et al.  The Drosophila pho-like gene encodes a YY1-related DNA binding protein that is redundant with pleiohomeotic in homeotic gene silencing , 2003, Development.

[31]  R. Urrutia,et al.  Sp1- and Krüppel-like transcription factors , 2003, Genome Biology.

[32]  Jürg Müller,et al.  The DNA-binding polycomb group protein pleiohomeotic mediates silencing of a Drosophila homeotic gene. , 1999, Development.

[33]  O. Cuvier,et al.  Recruitment of Drosophila Polycomb group proteins to chromatin by DSP1 , 2005, Nature.

[34]  D. Andrew,et al.  Epithelial Tube Morphology Is Determined by the Polarized Growth and Delivery of Apical Membrane , 2002, Cell.

[35]  S. Poux,et al.  Structure of a Polycomb Response Element and In Vitro Binding of Polycomb Group Complexes Containing GAGA Factor , 2000, Molecular and Cellular Biology.

[36]  A. Mohd-Sarip,et al.  Pleiohomeotic Can Link Polycomb to DNA and Mediate Transcriptional Repression , 2002, Molecular and Cellular Biology.

[37]  Yuh-Long Chang,et al.  Isolation and characterization of CHRASCH, a polycomb-containing silencing complex. , 2004, Methods in enzymology.

[38]  A. Birve,et al.  A 1-Megadalton ESC/E(Z) Complex from Drosophila That Contains Polycomblike and RPD3 , 2003, Molecular and Cellular Biology.

[39]  M. Lehmann.  Anything else but GAGA: a nonhistone protein complex reshapes chromatin structure. , 2004, Trends in genetics : TIG.

[40]  T. Mahmoudi,et al.  GAGA facilitates binding of Pleiohomeotic to a chromatinized Polycomb response element. , 2003, Nucleic acids research.

[41]  Y. Jan,et al.  A gain-of-function screen for genes that affect the development of the Drosophila adult external sensory organ. , 2000, Genetics.

[42]  M. Bienz,et al.  Long range repression conferring boundaries of Ultrabithorax expression in the Drosophila embryo. , 1991, The EMBO journal.

[43]  H. Jäckle,et al.  Sp1/egr-like zinc-finger protein required for endoderm specification and germ-layer formation in Drosophila , 1994, Nature.

[44]  Yi Zhang,et al.  Hierarchical recruitment of polycomb group silencing complexes. , 2004, Molecular cell.

[45]  J. M. Shields,et al.  Identification of the DNA sequence that interacts with the gut-enriched Krüppel-like factor. , 1998, Nucleic acids research.

[46]  V. Pirrotta,et al.  Drosophila Enhancer of Zeste/ESC Complexes Have a Histone H3 Methyltransferase Activity that Marks Chromosomal Polycomb Sites , 2002, Cell.

[47]  Giacomo Cavalli,et al.  Chromatin inheritance upon Zeste‐mediated Brahma recruitment at a minimal cellular memory module , 2004, The EMBO journal.

[48]  Jeng-Shin Lee,et al.  Evidence for physical interaction between the zinc-finger transcription factors YY1 and Sp1. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[49]  J. Hodgson,et al.  Site-Specific Recognition of a 70-Base-Pair Element Containing d(GA)n Repeats Mediatesbithoraxoid Polycomb Group Response Element-Dependent Silencing , 2001, Molecular and Cellular Biology.

[50]  V. Pirrotta,et al.  The mcp element from the Drosophila melanogaster bithorax complex mediates long-distance regulatory interactions. , 1999, Genetics.

[51]  M. Zavortink,et al.  The MCP silencer of the Drosophila Abd-B gene requires both Pleiohomeotic and GAGA factor for the maintenance of repression. , 2001, Development.

[52]  M. Bhat,et al.  Identification of the Drosophila progenitor of mammalian Krüppel-like factors 6 and 7 and a determinant of fly development. , 2003, Gene.

[53]  T. Steitz,et al.  Sequence-specific recognition of DNA by zinc-finger peptides derived from the transcription factor Sp1. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[54]  R. Kingston,et al.  A Drosophila Polycomb group complex includes Zeste and dTAFII proteins , 2001, Nature.

[55]  M. Fujioka,et al.  Analysis of an even-skipped rescue transgene reveals both composite and discrete neuronal and early blastoderm enhancers, and multi-stripe positioning by gap gene repressor gradients. , 1999, Development.