Abstract
The scabrous (sca) gene encodes a secreted dimeric glycoprotein with putative coiled-coil domains N-terminally and a C-terminal region related to the blood clot protein fibrinogen. Homozygous sca mutants have extra bristle organs and rough eyes. We describe a GAL4-based expression system for testing rescue of the sca mutant phenotype by altered SCA proteins and for misexpression. We find that deletion of the fibrinogen-related domain (FReD) greatly decreases SCA function, confirming the importance of this conserved region. SCA function could not be restored by FReDs from human fibrinogen chain genes. However, proteins lacking any FReD still showed some function in both rescue and misexpression experiments, suggesting that putative effector-binding regions lie outside this domain. Consistent with this, proteins expressing only the FReD had no rescuing activity but were recessive negative; i.e., they enhanced the phenotype of sca mutations but had no phenotype in the presence of a wild-type sca allele. This suggests that the FReD contributes to SCA function by binding to other components of the bristle determination pathway, increasing the activity of the linked N-terminal region.
MeSH Terms
Amino Acid Sequence
Animals
Base Sequence
Cloning, Molecular
Conserved Sequence
DNA-Binding Proteins
Drosophila Proteins
Drosophila melanogaster/genetics
Female
Fibrinogen/genetics
Fungal Proteins/genetics
Gene Expression
Genes, Insect/physiology
Glycoproteins/genetics,physiology
Humans
Insect Proteins/genetics,physiology
Male
Molecular Sequence Data
Mutagenesis, Insertional
Mutation
Recombinant Fusion Proteins
Saccharomyces cerevisiae Proteins
Sequence Deletion
Transcription Factors
Transgenes
Chemicals
DNA-Binding Proteins
Drosophila Proteins
Fungal Proteins
GAL4 protein, S cerevisiae
Glycoproteins
Insect Proteins
Recombinant Fusion Proteins
Saccharomyces cerevisiae Proteins
Transcription Factors
sca protein, Drosophila
Fibrinogen
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Lee E C
Department of Molecular Genetics, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
Yu S Y
Hu X
Mlodzik M
Baker N E
References (26)
26 references, click to expand
-
Characterization of complementary deoxyribonucleic acid and genomic deoxyribonucleic acid for the beta chain of human fibrinogen.
Biochemistry. 1983 Jun 21;22(13):3244-50
PMID: 6688356
-
Specificity of bone morphogenetic protein-related factors: cell fate and gene expression changes in Drosophila embryos induced by decapentaplegic but not 60A.
Cell Growth Differ. 1994 Jun;5(6):585-93
PMID: 8086336
-
Spacing differentiation in the developing Drosophila eye: a fibrinogen-related lateral inhibitor encoded by scabrous.
Science. 1990 Dec 7;250(4986):1370-7
PMID: 2175046
-
Isolation and expression of scabrous, a gene regulating neurogenesis in Drosophila.
Genes Dev. 1990 Nov;4(11):1848-61
PMID: 2125959
-
cDNA sequence of a second fibrinogen alpha chain in lamprey: an archetypal version alignable with full-length beta and gamma chains.
Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2066-70
PMID: 1549566
-
The role of fibrin in tumor metastasis.
Cancer Metastasis Rev. 1992 Nov;11(3-4):283-90
PMID: 1423819
-
Carboxy-terminal-extended variant of the human fibrinogen alpha subunit: a novel exon conferring marked homology to beta and gamma subunits.
Biochemistry. 1992 Dec 8;31(48):11968-72
PMID: 1457396
-
Confrontation of scabrous expressing and non-expressing cells is essential for normal ommatidial spacing in the Drosophila eye.
Development. 1994 Jul;120(7):1959-69
PMID: 7925001
-
Fibrinogen in human atherosclerosis.
Ann N Y Acad Sci. 1995 Jan 17;748:461-71; DISCUSSION 471-3
PMID: 7695189
-
Notch signaling.
Science. 1995 Apr 14;268(5208):225-32
PMID: 7716513
-
Drosophila eye development: Notch and Delta amplify a neurogenic pattern conferred on the morphogenetic furrow by scabrous.
Mech Dev. 1995 Feb;49(3):173-89
PMID: 7734391
-
Resolution of spontaneous bleeding events but failure of pregnancy in fibrinogen-deficient mice.
Genes Dev. 1995 Aug 15;9(16):2020-33
PMID: 7649481
-
Molecular analysis of scabrous mutant alleles from Drosophila melanogaster indicates a secreted protein with two functional domains.
Genetics. 1995 Oct;141(2):607-17
PMID: 8647397
-
The scabrous gene encodes a secreted glycoprotein dimer and regulates proneural development in Drosophila eyes.
Mol Cell Biol. 1996 Mar;16(3):1179-88
PMID: 8622662
-
gp300sca is not a high affinity Notch ligand.
Biochem Biophys Res Commun. 1996 Aug 23;225(3):720-5
PMID: 8780680
-
Isolation of angiopoietin-1, a ligand for the TIE2 receptor, by secretion-trap expression cloning.
Cell. 1996 Dec 27;87(7):1161-9
PMID: 8980223
-
Requisite role of angiopoietin-1, a ligand for the TIE2 receptor, during embryonic angiogenesis.
Cell. 1996 Dec 27;87(7):1171-80
PMID: 8980224
-
Angiopoietin-2, a natural antagonist for Tie2 that disrupts in vivo angiogenesis.
Science. 1997 Jul 4;277(5322):55-60
PMID: 9204896
-
The primary fibrin polymerization pocket: three-dimensional structure of a 30-kDa C-terminal gamma chain fragment complexed with the peptide Gly-Pro-Arg-Pro.
Proc Natl Acad Sci U S A. 1997 Jul 8;94(14):7176-81
PMID: 9207064
-
Crystal structures of fragment D from human fibrinogen and its crosslinked counterpart from fibrin.
Nature. 1997 Oct 2;389(6650):455-62
PMID: 9333233
-
Preferential transposition of Drosophila P elements to nearby chromosomal sites.
Genetics. 1993 Feb;133(2):347-59
PMID: 8382177
-
A detailed consideration of a principal domain of vertebrate fibrinogen and its relatives.
Protein Sci. 1992 Dec;1(12):1563-77
PMID: 1304888
-
Fibrinogen mediates leukocyte adhesion to vascular endothelium through an ICAM-1-dependent pathway.
Cell. 1993 Jul 2;73(7):1423-34
PMID: 8100742
-
Targeted gene expression as a means of altering cell fates and generating dominant phenotypes.
Development. 1993 Jun;118(2):401-15
PMID: 8223268
-
Tenascin-C, tenascin-R and tenascin-X: a family of talented proteins in search of functions.
Curr Opin Cell Biol. 1993 Oct;5(5):869-76
PMID: 7694605
-
Tumors: wounds that do not heal. Similarities between tumor stroma generation and wound healing.
N Engl J Med. 1986 Dec 25;315(26):1650-9
PMID: 3537791