Mechanism of Regulatory Target Selection by the SOX High-Mobility-Group Domain Proteins as Revealed by Comparison of SOX1/2/3 and SOX9

ABSTRACT SOX proteins bind similar DNA motifs through their high-mobility-group (HMG) domains, but their action is highly specific with respect to target genes and cell type. We investigated the mechanism of target selection by comparing SOX1/2/3, which activate δ-crystallin minimal enhancer DC5, with SOX9, which activates Col2a1 minimal enhancer COL2C2. These enhancers depend on both the SOX binding site and the binding site of a putative partner factor. The DC5 site was equally bound and bent by the HMG domains of SOX1/2 and SOX9. The activation domains of these SOX proteins mapped at the distal portions of the C-terminal domains were not cell specific and were independent of the partner factor. Chimeric proteins produced between SOX1 and SOX9 showed that to activate the DC5 enhancer, the C-terminal domain must be that of SOX1, although the HMG domains were replaceable. The SOX2-VP16 fusion protein, in which the activation domain of SOX2 was replaced by that of VP16, activated the DC5 enhancer still in a partner factor-dependent manner. The results argue that the proximal portion of the C-terminal domain of SOX1/2 specifically interacts with the partner factor, and this interaction determines the specificity of the SOX1/2 action. Essentially the same results were obtained in the converse experiments in which COL2C2 activation by SOX9 was analyzed, except that specificity of SOX9-partner factor interaction also involved the SOX9 HMG domain. The highly selective SOX-partner factor interactions presumably stabilize the DNA binding of the SOX proteins and provide the mechanism for regulatory target selection.

[1]  H. Okayama,et al.  High-efficiency transformation of mammalian cells by plasmid DNA. , 1987, Molecular and cellular biology.

[2]  K. Goto,et al.  Lens-specific enhancer in the third intron regulates expression of the chicken delta 1-crystallin gene. , 1987, Genes & development.

[3]  Jun Ma,et al.  GAL4-VP16 is an unusually potent transcriptional activator , 1988, Nature.

[4]  C. Zwieb,et al.  Bending of DNA by gene-regulatory proteins: construction and use of a DNA bending vector. , 1989, Gene.

[5]  J. Leatherwood,et al.  An amino-terminal fragment of GAL4 binds DNA as a dimer. , 1989, Journal of molecular biology.

[6]  N. Nakatsuji,et al.  A mouse embryonic stem cell line showing pluripotency of differentiation in early embryos and ubiquitous beta-galactosidase expression. , 1990, Cell differentiation and development : the official journal of the International Society of Developmental Biologists.

[7]  M. Scott,et al.  What determines the specificity of action of Drosophila homeodomain proteins? , 1990, Cell.

[8]  Peter Goodfellow,et al.  A gene mapping to the sex-determining region of the mouse Y chromosome is a member of a novel family of embryonically expressed genes , 1990, Nature.

[9]  G. Nolan,et al.  Improved FACS-Gal: flow cytometric analysis and sorting of viable eukaryotic cells expressing reporter gene constructs. , 1991, Cytometry.

[10]  R Grosschedl,et al.  DNA-binding properties of the HMG domain of the lymphoid-specific transcriptional regulator LEF-1. , 1991, Genes & development.

[11]  A. Ashworth,et al.  An SRY‐related gene expressed during spermatogenesis in the mouse encodes a sequence‐specific DNA‐binding protein. , 1992, The EMBO journal.

[12]  P N Goodfellow,et al.  SRY, like HMG1, recognizes sharp angles in DNA. , 1992, The EMBO journal.

[13]  A. Ashworth,et al.  A conserved family of genes related to the testis determining gene, SRY. , 1992, Nucleic acids research.

[14]  K. Jones,et al.  The hLEF/TCF-1 alpha HMG protein contains a context-dependent transcriptional activation domain that induces the TCR alpha enhancer in T cells. , 1993, Genes & development.

[15]  H. Clevers,et al.  Sox‐4, an Sry‐like HMG box protein, is a transcriptional activator in lymphocytes. , 1993, The EMBO journal.

[16]  H. Kondoh,et al.  Overlapping positive and negative regulatory elements determine lens-specific activity of the delta 1-crystallin enhancer , 1993, Molecular and cellular biology.

[17]  P. Koopman,et al.  Seven new members of the Sox gene family expressed during mouse development. , 1993, Nucleic acids research.

[18]  R. Grosschedl,et al.  LEF‐1 contains an activation domain that stimulates transcription only in a specific context of factor‐binding sites. , 1993, The EMBO journal.

[19]  P. Goodfellow,et al.  Definition of a consensus DNA binding site for SRY. , 1994, Nucleic acids research.

[20]  R Grosschedl,et al.  Distinct DNA-binding properties of the high mobility group domain of murine and human SRY sex-determining factors. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[21]  R. Lovell-Badge,et al.  Sex‐reversing mutations affect the architecture of SRY‐DNA complexes. , 1994, The EMBO journal.

[22]  N. Tommerup,et al.  Autosomal sex reversal and campomelic dysplasia are caused by mutations in and around the SRY-related gene SOX9 , 1994, Cell.

[23]  A. Ashworth,et al.  DNA binding and bending properties of the post-meiotically expressed Sry-related protein Sox-5. , 1994, Nucleic acids research.

[24]  Andy Greenfield,et al.  The Sry-related gene Sox9 is expressed during chondrogenesis in mouse embryos , 1995, Nature Genetics.

[25]  R. Lovell-Badge,et al.  Involvement of SOX proteins in lens‐specific activation of crystallin genes. , 1995, The EMBO journal.

[26]  N. Corbi,et al.  Developmental-specific activity of the FGF-4 enhancer requires the synergistic action of Sox2 and Oct-3. , 1995, Genes & development.

[27]  A. Ashworth,et al.  The Sry-related HMG box-containing gene Sox6 is expressed in the adult testis and developing nervous system of the mouse. , 1995, Nucleic acids research.

[28]  E. Cartwright,et al.  Embryonic expression of the chicken Sox2, Sox3 and Sox11 genes suggests an interactive role in neuronal development , 1995, Mechanisms of Development.

[29]  R. Mann,et al.  Extra specificity from extradenticle: the partnership between HOX and PBX/EXD homeodomain proteins. , 1996, Trends in genetics : TIG.

[30]  V. Lefebvre,et al.  An 18-base-pair sequence in the mouse proalpha1(II) collagen gene is sufficient for expression in cartilage and binds nuclear proteins that are selectively expressed in chondrocytes , 1996, Molecular and cellular biology.

[31]  P. Goodfellow,et al.  A comparison of the properties of Sox-3 with Sry and two related genes, Sox-1 and Sox-2. , 1996, Development.

[32]  Y. Kanai,et al.  Identification of two Sox17 messenger RNA isoforms, with and without the high mobility group box region, and their differential expression in mouse spermatogenesis , 1996, The Journal of cell biology.

[33]  P H Krebsbach,et al.  Identification of a Minimum Enhancer Sequence for the Type II Collagen Gene Reveals Several Core Sequence Motifs in Common with the Link Protein Gene (*) , 1996, The Journal of Biological Chemistry.

[34]  A. Sinclair,et al.  A male-specific role for SOX9 in vertebrate sex determination. , 1996, Development.

[35]  Gerd Scherer,et al.  Sex reversal by loss of the C–terminal transactivation domain of human SOX9 , 1996, Nature Genetics.

[36]  P. Goodfellow,et al.  Sox9 expression during gonadal development implies a conserved role for the gene in testis differentiation in mammals and birds , 1996, Nature Genetics.

[37]  J. Scholey.  Kinesin-II, a membrane traffic motor in axons, axonemes, and spindles , 1996, The Journal of cell biology.

[38]  R. Lovell-Badge,et al.  Sox genes find their feet. , 1997, Current opinion in genetics & development.

[39]  R. Lovell-Badge,et al.  Preventing the loss of competence for neural induction: HGF/SF, L5 and Sox-2. , 1997, Development.

[40]  D. Ambrosetti,et al.  Synergistic activation of the fibroblast growth factor 4 enhancer by Sox2 and Oct-3 depends on protein-protein interactions facilitated by a specific spatial arrangement of factor binding sites , 1997, Molecular and cellular biology.

[41]  P. Koopman,et al.  Expression of the Sox11 gene in mouse embryos suggests roles in neuronal maturation and epithelio‐mesenchymal induction , 1997, Developmental dynamics : an official publication of the American Association of Anatomists.

[42]  H. Clevers,et al.  Cell differentiation by interaction of two HMG‐box proteins: Mat1‐Mc activates M cell‐specific genes in S.pombe by recruiting the ubiquitous transcription factor Ste11 to weak binding sites , 1997, The EMBO journal.

[43]  V. Lefebvre,et al.  Parallel expression of Sox9 and Col2a1 in cells undergoing chondrogenesis , 1997, Developmental dynamics : an official publication of the American Association of Anatomists.

[44]  P. Sharpe,et al.  Dynamic expression of chicken Sox2 and Sox3 genes in ectoderm induced to form neural tissue , 1997, Developmental dynamics : an official publication of the American Association of Anatomists.

[45]  Donald M. Bell,et al.  SOX9 binds DNA, activates transcription, and coexpresses with type II collagen during chondrogenesis in the mouse. , 1997, Developmental biology.

[46]  P N Goodfellow,et al.  SOX9 is a potent activator of the chondrocyte-specific enhancer of the pro alpha1(II) collagen gene , 1997, Molecular and cellular biology.

[47]  Patrick P.L. Tam,et al.  SOX9 directly regulates the type-ll collagen gene , 1997, Nature Genetics.

[48]  R. Lovell-Badge,et al.  Involvement of Sox1, 2 and 3 in the early and subsequent molecular events of lens induction. , 1998, Development.

[49]  R. Lovell-Badge,et al.  Sox1 directly regulates the gamma-crystallin genes and is essential for lens development in mice. , 1998, Genes & development.

[50]  R. Lovell-Badge,et al.  A role for SOX1 in neural determination. , 1998, Development.