title: A Sox10 expression screen identifies an amino acid essential for Erbb3 function.
abstract: The neural crest (NC) is a population of embryonic stem cells that gives rise to numerous cell types, including the glia and neurons of the peripheral and enteric nervous systems and the melanocytes of the skin and hair. Mutations in genes and genetic pathways regulating NC development lead to a wide spectrum of human developmental disorders collectively called neurocristopathies. To identify molecular pathways regulating NC development and to understand how alterations in these processes lead to disease, we established an N-ethyl-N-nitrosourea (ENU) mutagenesis screen utilizing a mouse model sensitized for NC defects, Sox10(LacZ/+). Out of 71 pedigrees analyzed, we identified and mapped four heritable loci, called modifier of Sox10 expression pattern 1-4 (msp1-4), which show altered NC patterning. In homozygous msp1 embryos, Sox10(LacZ) expression is absent in cranial ganglia, cranial nerves, and the sympathetic chain; however, the development of other Sox10-expressing cells appears unaffected by the mutation. Linkage analysis, sequencing, and complementation testing confirmed that msp1 is a new allele of the receptor tyrosine kinase Erbb3, Erbb3(msp1), that carries a single amino acid substitution in the extracellular region of the protein. The ENU-induced mutation does not alter protein expression, however, it is sufficient to impair ERBB3 signaling such that the embryonic defects observed in msp1 resemble those of Erbb3 null alleles. Biochemical analysis of the mutant protein showed that ERBB3 is expressed on the cell surface, but its ligand-induced phosphorylation is dramatically reduced by the msp1 mutation. These findings highlight the importance of the mutated residue for ERBB3 receptor function and activation. This study underscores the utility of using an ENU mutagenesis to identify genetic pathways regulating NC development and to dissect the roles of discrete protein domains, both of which contribute to a better understanding of gene function in a cellular and developmental setting.
authors.affiliation: Genetic Disease Research Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland, United States of America.
authors.forename: Kristina
authors.initials: K
authors.lastname: Buac
authors.forename: Dawn E
authors.initials: DE
authors.lastname: Watkins-Chow
authors.forename: Stacie K
authors.initials: SK
authors.lastname: Loftus
authors.forename: Denise M
authors.initials: DM
authors.lastname: Larson
authors.forename: Arturo
authors.initials: A
authors.lastname: Incao
authors.forename: Gretchen
authors.initials: G
authors.lastname: Gibney
authors.forename: William J
authors.initials: WJ
authors.lastname: Pavan
mesh_terms: D000483:Alleles; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000596:Amino Acids / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D005038:Ethylnitrosourea / Q000494:pharmacology; D015870:Gene Expression; D006609:High Mobility Group Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009154:Mutation; D009432:Neural Crest / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D010641:Phenotype; D010766:Phosphorylation; D012333:RNA, Messenger / Q000378:metabolism; D020893:Receptor, ErbB-3 / Q000235:genetics* / Q000378:metabolism*; D055757:SOXE Transcription Factors; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism
chemical_list: D000596:Amino Acids; D004268:DNA-Binding Proteins; D006609:High Mobility Group Proteins; D012333:RNA, Messenger; C527923:SOX10 protein, human; D055757:SOXE Transcription Factors; C527924:Sox10 protein, mouse; D014157:Transcription Factors; D020893:Receptor, ErbB-3; D005038:Ethylnitrosourea
references.citation: Proc Natl Acad Sci U S A. 1998 Jun 23;95(13):7485-90
references.pmid: 9636176
references.citation: PLoS Biol. 2004 Aug;2(8):E219
references.pmid: 15314648
references.citation: Nat Genet. 2002 Feb;30(2):185-9
references.pmid: 11818962
references.citation: J Neurosci. 1998 Jan 1;18(1):237-50
references.pmid: 9412504
references.citation: Development. 2000 Dec;127(24):5379-89
references.pmid: 11076759
references.citation: Hum Mol Genet. 1999 Sep;8(9):1785-9
references.pmid: 10441344
references.citation: Proc Natl Acad Sci U S A. 1990 Jul;87(13):4905-9
references.pmid: 2164210
references.citation: Nat Genet. 1998 Feb;18(2):171-3
references.pmid: 9462749
references.citation: J Biol Chem. 1994 May 13;269(19):14303-6
references.pmid: 8188716
references.citation: Mech Dev. 2005 May;122(5):659-69
references.pmid: 15817223
references.citation: Nat Genet. 1998 Jan;18(1):60-4
references.pmid: 9425902
references.citation: J Biol Chem. 1998 Jun 26;273(26):16050-7
references.pmid: 9632656
references.citation: J Biol. 2005;4(3):10
references.pmid: 16492316
references.citation: Mol Cell Biol. 2005 Sep;25(17):7734-42
references.pmid: 16107719
references.citation: Hum Mol Genet. 2007 Dec 15;16(24):3037-46
references.pmid: 17855451
references.citation: Mol Biol Cell. 2003 Jul;14(7):2756-67
references.pmid: 12857862
references.citation: Neuron. 2005 Feb 17;45(4):513-23
references.pmid: 15721238
references.citation: Development. 2003 Jun;130(12):2809-18
references.pmid: 12736222
references.citation: J Biol Chem. 2008 Mar 28;283(13):8229-36
references.pmid: 18216017
references.citation: Cell. 2002 Sep 20;110(6):775-87
references.pmid: 12297050
references.citation: Proc Natl Acad Sci U S A. 2005 Apr 26;102(17):5913-9
references.pmid: 15755804
references.citation: Cell. 2002 Sep 20;110(6):763-73
references.pmid: 12297049
references.citation: Science. 2002 Aug 23;297(5585):1330-3
references.pmid: 12154198
references.citation: Proc Natl Acad Sci U S A. 1994 Aug 16;91(17):8132-6
references.pmid: 8058768
references.citation: Genes Dev. 2001 Jan 1;15(1):66-78
references.pmid: 11156606
references.citation: J Med Genet. 2001 Sep;38(9):E30
references.pmid: 11546831
references.citation: Am J Hum Genet. 2000 May;66(5):1496-503
references.pmid: 10762540
references.citation: Int Rev Cytol. 1986;103:89-145
references.pmid: 3528022
references.citation: J Biol Chem. 1994 May 20;269(20):14661-5
references.pmid: 7514177
references.citation: Hum Genet. 2000 Jul;107(1):1-6
references.pmid: 10982026
references.citation: Nucleic Acids Res. 1999 Mar 15;27(6):1409-20
references.pmid: 10037800
references.citation: Nature. 1995 Nov 23;378(6555):386-90
references.pmid: 7477375
references.citation: Development. 1999 Apr;126(8):1601-9
references.pmid: 10079223
references.citation: Nature. 1997 Oct 16;389(6652):725-30
references.pmid: 9338783
references.citation: Nature. 1991 Dec 19-26;354(6354):526-8
references.pmid: 1722027
references.citation: Dev Biol. 1987 May;121(1):220-36
references.pmid: 3552788
references.citation: Hum Mol Genet. 2008 Jul 15;17(14):2118-31
references.pmid: 18397875
references.citation: Neuron. 2003 Apr 10;38(1):17-31
references.pmid: 12691661
references.citation: Mamm Genome. 1998 Feb;9(2):150-4
references.pmid: 9457677
references.citation: Proteins. 1995 Nov;23(3):318-26
references.pmid: 8710825
references.citation: Development. 1997 Sep;124(18):3575-86
references.pmid: 9342050
references.citation: Proc Natl Acad Sci U S A. 2002 Apr 2;99(7):4471-6
references.pmid: 11917121
references.citation: Development. 1997 Dec;124(24):4999-5011
references.pmid: 9362461