Cysteine Repeat Domains and Adjacent Sequences Determine Distinct Bone Morphogenetic Protein Modulatory Activities of the Drosophila Sog Protein

The Drosophila short gastrulation gene (sog) encodes a large extracellular protein (Sog) that inhibits signaling by BMP-related ligands. Sog and its vertebrate counterpart Chordin contain four copies of a cysteine repeat (CR) motif defined by 10 cysteine residues spaced in a fixed pattern and a tryptophan residue situated between the first two cysteines. Here we present a structure-function analysis of the CR repeats in Sog, using a series of deletion and point mutation constructs, as well as constructs in which CR domains have been swapped. This analysis indicates that the CR domains are individually dispensable for Sog function but that they are not interchangeable. These studies reveal three different types of Sog activity: intact Sog, which inhibits signaling mediated by the ligand Glass bottom boat (Gbb), a more broadly active class of BMP antagonist referred to as Supersog, and a newly identified activity, which may promote rather than inhibit BMP signaling. Analysis of the activities of CR swap constructs indicates that the CR domains are required for full activity of the various forms of Sog but that the type of Sog activity is determined primarily by surrounding protein sequences. Cumulatively, our analysis suggests that CR domains interact physically with adjacent protein sequences to create forms of Sog with distinct BMP modulatory activities.

[1]  O. Shimmi,et al.  Physical properties of Tld, Sog, Tsg and Dpp protein interactions are predicted to help create a sharp boundary in Bmp signals during dorsoventral patterning of the Drosophila embryo , 2003, Development.

[2]  CHRD, a novel domain in the BMP inhibitor chordin, is also found in microbial proteins. , 2003, Trends in biochemical sciences.

[3]  W. Van Hul,et al.  Extracellular regulation of BMP signaling in vertebrates: a cocktail of modulators. , 2002, Developmental biology.

[4]  N. Barkai,et al.  Robustness of the BMP morphogen gradient in Drosophila embryonic patterning , 2022 .

[5]  K. Cadigan.  Regulating morphogen gradients in the Drosophila wing. , 2002, Seminars in cell & developmental biology.

[6]  R. Ray,et al.  Context-dependent relationships between the BMPs gbb and dpp during development of the Drosophila wing imaginal disk. , 2001, Development.

[7]  E. L. Ferguson,et al.  A positive role for Short gastrulation in modulating BMP signaling during dorsoventral patterning in the Drosophila embryo. , 2001, Development.

[8]  P. Lemaire,et al.  The human chordin gene encodes several differentially expressed spliced variants with distinct BMP opposing activities , 2001, Mechanisms of Development.

[9]  A. Brivanlou,et al.  Twisted gastrulation can function as a BMP antagonist , 2001, Nature.

[10]  Ken W. Y. Cho,et al.  Homologues of Twisted gastrulation are extracellular cofactors in antagonism of BMP signalling , 2001, Nature.

[11]  Stephen C. Ekker,et al.  Twisted gastrulation is a conserved extracellular BMP antagonist , 2001, Nature.

[12]  D. J. Olson,et al.  Crossveinless 2 contains cysteine-rich domains and is required for high levels of BMP-like activity during the formation of the cross veins in Drosophila. , 2000, Development.

[13]  E. Robertis,et al.  The evolutionarily conserved BMP-binding protein Twisted gastrulation promotes BMP signalling , 2000, Nature.

[14]  O. Shimmi,et al.  Processing of the Drosophila Sog protein creates a novel BMP inhibitory activity. , 2000, Development.

[15]  N. Dostatni,et al.  Persistence of Hunchback in the terminal region of the Drosophila blastoderm embryo impairs anterior development. , 2000, Development.

[16]  J. Massagué,et al.  Controlling TGF-β signaling , 2000, Genes & Development.

[17]  E. D. De Robertis,et al.  BMP-binding modules in chordin: a model for signalling regulation in the extracellular space. , 2000, Development.

[18]  Michael Levine,et al.  Local inhibition and long-range enhancement of Dpp signal transduction by Sog , 1999, Nature.

[19]  Sangbin Park,et al.  Interpretation of a BMP Activity Gradient in Drosophila Embryos Depends on Synergistic Signaling by Two Type I Receptors, SAX and TKV , 1998, Cell.

[20]  E. L. Ferguson,et al.  Spatially Restricted Activation of the SAX Receptor by SCW Modulates DPP/TKV Signaling in Drosophila Dorsal–Ventral Patterning , 1998, Cell.

[21]  B. Biehs,et al.  The knirps and knirps-related genes organize development of the second wing vein in Drosophila. , 1998, Development.

[22]  M. O’Connor,et al.  Synergistic signaling by two BMP ligands through the SAX and TKV receptors controls wing growth and patterning in Drosophila. , 1998, Development.

[23]  S. Torres-Schumann,et al.  TGF-beta/BMP superfamily members, Gbb-60A and Dpp, cooperate to provide pattern information and establish cell identity in the Drosophila wing. , 1998, Development.

[24]  Leslie Dale,et al.  Cleavage of Chordin by Xolloid Metalloprotease Suggests a Role for Proteolytic Processing in the Regulation of Spemann Organizer Activity , 1997, Cell.

[25]  Ken W. Y. Cho,et al.  Production of a DPP Activity Gradient in the Early Drosophila Embryo through the Opposing Actions of the SOG and TLD Proteins , 1997, Cell.

[26]  E. Bier.  Anti-Neural-Inhibition: A Conserved Mechanism for Neural Induction , 1997, Cell.

[27]  D. Melton,et al.  Vertebrate Embryonic Cells Will Become Nerve Cells Unless Told Otherwise , 1997, Cell.

[28]  B. Biehs,et al.  The Drosophila decapentaplegic and short gastrulation genes function antagonistically during adult wing vein development. , 1996, Development.

[29]  B. Biehs,et al.  The Drosophila short gastrulation gene prevents Dpp from autoactivating and suppressing neurogenesis in the neuroectoderm. , 1996, Genes & development.

[30]  A. McMahon,et al.  Genetic analysis of dorsoventral pattern formation in the zebrafish: requirement of a BMP-like ventralizing activity and its dorsal repressor. , 1996, Genes & development.

[31]  Y. Sasai,et al.  Dorsoventral Patterning in Xenopus: Inhibition of Ventral Signals by Direct Binding of Chordin to BMP-4 , 1996, Cell.

[32]  D. Kimelman,et al.  Drosophila short gastrulation induces an ectopic axis in Xenopus: evidence for conserved mechanisms of dorsal-ventral patterning. , 1995, Development.

[33]  P. Wilson,et al.  Induction of epidermis and inhibition of neural fate by Bmp-4 , 1995, Nature.

[34]  Yoshiki Sasai,et al.  A conserved system for dorsal-ventral patterning in insects and vertebrates involving sog and chordin , 1995, Nature.

[35]  E. Bier,et al.  Xenopus chordin and Drosophila short gastrulation genes encode homologous proteins functioning in dorsal-ventral axis formation , 1995, Cell.

[36]  Y. Sasai,et al.  Regulation of neural induction by the Chd and Bmp-4 antagonistic patterning signals in Xenopus , 1995, Nature.

[37]  Y. Sasai,et al.  Xenopus chordin: A novel dorsalizing factor activated by organizer-specific homeobox genes , 1994, Cell.

[38]  J. Emery,et al.  Dorsal-ventral patterning of the Drosophila embryo depends on a putative negative growth factor encoded by the short gastrulation gene. , 1994, Genes & development.

[39]  E. Bier,et al.  Double-label in situ hybridization using biotin and digoxigenin-tagged RNA probes. , 1994, BioTechniques.

[40]  M. Levine,et al.  The screw gene encodes a ubiquitously expressed member of the TGF-beta family required for specification of dorsal cell fates in the Drosophila embryo. , 1994, Genes & development.

[41]  I. Guerrero,et al.  Targeted expression of the signaling molecule decapentaplegic induces pattern duplications and growth alterations in Drosophila wings. , 1994, The EMBO journal.

[42]  E. Bier,et al.  New functions of the Drosophila rhomboid gene during embryonic and adult development are revealed by a novel genetic method, enhancer piracy. , 1994, Development.

[43]  J. Marsh,et al.  Dorsal midline fate in Drosophila embryos requires twisted gastrulation, a gene encoding a secreted protein related to human connective tissue growth factor. , 1994, Genes & development.

[44]  H. Weintraub,et al.  Expression of achaete-scute homolog 3 in Xenopus embryos converts ectodermal cells to a neural fate. , 1994, Genes & development.

[45]  E. Bier,et al.  The Drosophila rhomboid gene mediates the localized formation of wing veins and interacts genetically with components of the EGF-R signaling pathway. , 1993, Genes & development.

[46]  N. Perrimon,et al.  Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. , 1993, Development.

[47]  W. Gelbart,et al.  Human BMP sequences can confer normal dorsal-ventral patterning in the Drosophila embryo. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[48]  W. Gelbart,et al.  An activity gradient of decapentaplegic is necessary for the specification of dorsal pattern elements in the Drosophila embryo. , 1993, Development.

[49]  J. Massagué,et al.  The transforming growth factor-beta family. , 1990, Annual review of cell biology.

[50]  W. Gelbart,et al.  A transcript from a Drosophila pattern gene predicts a protein homologous to the transforming growth factor-β family , 1987, Nature.

[51]  W. Gelbart,et al.  Shortvein, a new component of the decapentaplegic gene complex in Drosophila melanogaster. , 1985, Genetics.