A screen for dominant mutations applied to components in the Drosophila EGF-R pathway.
The Drosophila epidermal growth factor receptor (EGF-R) controls many critical cell fate choices throughout development. Several proteins collaborate to promote localized EGF-R activation, such as Star and Rhomboid (Rho), which act sequentially to ensure the maturation and processing of inactive membrane-bound EGF ligands. To gain insights into the mechanisms underlying Rho and Star function, we developed a mutagenesis scheme to isolate novel overexpression activity (NOVA) alleles. In the case of rho, we isolated a dominant neomorphic allele, which interferes with Notch signaling, as well as a dominant-negative allele, which produces RNA interference-like flip-back transcripts that reduce endogenous rho expression. We also obtained dominant-negative and neomorphic Star mutations, which have phenotypes similar to those of rho NOVA alleles, as well as dominant-negative Egf-r alleles. The isolation of dominant alleles in several different genes suggests that NOVA mutagenesis should be widely applicable and emerge as an effective tool for generating dominant mutations in genes of unknown function.
Guichard
Annabel
A
Section of Cell and Developmental Biology, University of California at San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0349, USA.
Srinivasan
Shaila
S
Zimm
Georgianna
G
Bier
Ethan
E
D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism; D004331:Drosophila melanogaster / Q000196:embryology / Q000235:genetics / Q000378:metabolism*; D066246:ErbB Receptors / Q000378:metabolism*; D005260:Female; D015870:Gene Expression; D005799:Genes, Dominant / Q000235:genetics*; D017344:Genes, Insect / Q000235:genetics; D008297:Male; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000235:genetics*; D010641:Phenotype; D012313:RNA / Q000235:genetics / Q000378:metabolism; D051880:Receptors, Notch; D015398:Signal Transduction; D014921:Wings, Animal / Q000196:embryology / Q000502:physiology
D029721:Drosophila Proteins; D008565:Membrane Proteins; C497759:N protein, Drosophila; D051880:Receptors, Notch; C081481:Rho protein, Drosophila; C415248:S protein, Drosophila; C472498:Stet protein, Drosophila; D012313:RNA; D066246:ErbB Receptors