Functions of the cytoplasmic domain of the beta PS integrin subunit during Drosophila development.
Integrins constitute a family of membrane-spanning, heterodimeric proteins that mediate adhesive interactions between cells and surrounding extracellular matrices (or other cells) and participate in signal transduction. We are interested in assessing integrin functions in the context of developing Drosophila melanogaster. This report, using mutants of the beta PS subunit encoded by the myospheroid (mys) locus, analyzes the relationships between integrin protein structure and developmental functions in an intact organism. As a first step in this analysis, we demonstrated the ability of a fragment of wild-type mys genomic DNA, introduced into the germ line in a P-element vector P[mys+], to rescue phenotypes attributed to lack of (or defects in) the endogenous beta PS during several discrete morphogenetic events. We then produced in vitro a series of modifications of the wild-type P[mys+] transposon, which encode beta PS derivatives with mutations within the small and highly conserved cytoplasmic domain. In vivo analysis of these mutant transposons led to the following conclusions. (1) The cytoplasmic tail of beta PS is essential for all developmental functions of the protein that were assayed. (2) An intron at a conserved position in the DNA sequence encoding the cytoplasmic tail is thought to participate in important alternative splicing events in vertebrate beta integrin subunit genes, but is not required for the developmental functions of the mys gene assayed here. (3) Phosphorylation on two conserved tyrosines found in the C terminus of the beta PS cytoplasmic tail is not necessary for the tested developmental functions. (4) Four highly conserved amino acid residues found in the N-terminal portion of the cytoplasmic tail are important but not critical for the developmental functions of beta PS; furthermore, the efficiencies with which these mutant proteins function during different morphogenetic processes vary greatly, strongly suggesting that the cytoplasmic interactions involving PS integrins are developmentally modulated.
Grinblat
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Y
Department of Cellular and Developmental Biology, Harvard University, Cambridge, MA 02138.
Zusman
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Yee
G
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Hynes
R O
RO
Kafatos
F C
FC
D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion / Q000235:genetics; D003593:Cytoplasm / Q000502:physiology*; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000033:anatomy & histology / Q000196:embryology* / Q000235:genetics; D005821:Genetic Techniques; D039001:Integrin alpha Chains; D016023:Integrins / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D009024:Morphogenesis / Q000235:genetics; D009154:Mutation / Q000235:genetics; D010641:Phenotype
D029721:Drosophila Proteins; D039001:Integrin alpha Chains; D016023:Integrins; C082188:if protein, Drosophila