Identification of integrin beta subunit mutations that alter heterodimer function in situ.
We conducted a genetic screen for mutations in myospheroid, the gene encoding the Drosophila betaPS integrin subunit, and identified point mutants in all of the structural domains of the protein. Surprisingly, we find that mutations in very strongly conserved residues will often allow sufficient integrin function to support the development of adult animals, including mutations in the ADMIDAS site and in a cytoplasmic NPXY motif. Many mutations in the I-like domain reduce integrin expression specifically when betaPS is combined with activating alphaPS2 cytoplasmic mutations, indicating that integrins in the extended conformation are unstable relative to the inactive, bent heterodimers. Interestingly, the screen has identified alleles that show gain-of-function characteristics in cell culture, but have negative effects on animal development or viability. This is illustrated by the allele mys(b58); available structural models suggest that the molecular lesion of mys(b58), V409>D, should promote the "open" conformation of the beta subunit I-like domain. This expectation is supported by the finding that alphaPS2betaPS (V409>D) promotes adhesion and spreading of S2 cells more effectively than does wild-type alphaPS2betaPS, even when betaPS is paired with alphaPS2 containing activating cytoplasmic mutations. Finally, comparisons with the sequence of human beta8 suggest that evolution has targeted the "mys(b58)" residue as a means of affecting integrin activity.
Jannuzi
Alison L
AL
Department of Molecular and Cellular Biology, Arizona Cancer Center, University of Arizona, Tucson, Arizona 85724, USA.
Bunch
Thomas A
TA
West
Robert F
RF
Brower
Danny L
DL
D000483:Alleles; D020816:Amino Acid Motifs / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion / Q000235:genetics; D002465:Cell Movement / Q000235:genetics; D004330:Drosophila / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D029721:Drosophila Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004848:Epithelium / Q000737:chemistry; D005820:Genetic Testing; D039001:Integrin alpha Chains; D039641:Integrin beta Chains / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D016023:Integrins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D017354:Point Mutation; D017434:Protein Structure, Tertiary
D029721:Drosophila Proteins; D039001:Integrin alpha Chains; D039641:Integrin beta Chains; D016023:Integrins; C082188:if protein, Drosophila