Antineurogenic phenotypes induced by truncated Notch proteins indicate a role in signal transduction and may point to a novel function for Notch in nuclei.
Loss of any one of several neurogenic genes of Drosophila results in overproduction of embryonic neuroblasts at the expense of epidermoblasts. In this paper a variety of altered Notch proteins are expressed in transgenic flies. Dominant lethal, antineurogenic phenotypes were produced by expression of three classes of mutant proteins: (1) a protein comprised of the cytoplasmic domain of Notch and devoid of sequences permitting membrane association; (2) a transmembrane protein lacking the extracellular, lin12/Notch repeats; and (3) transmembrane proteins carrying amino acid substitutions replacing one or both extracellular cysteines thought to be involved in Notch dimerization. These Notch proteins not only suppress the neural hypertrophy observed in Notch- embryos, but also generate a phenotype in which elements of the embryonic nervous system are underproduced. Action of the intracellular cdc10 repeats appears to be essential for wild-type Notch function or for the antineurogenic activity of these proteins. The activities of the dominant, gain-of-function proteins indicate that Notch functions as a signal transducing receptor during ectoderm development. Production of antineurogenic Notch proteins in embryos deficient for the other neurogenic genes allowed functional dependencies to be established. Delta, mastermind, bigbrain, and neuralized appear to function in elaboration of a signal upstream of Notch. Genes of the Enhancer of split complex act after Notch. The cytoplasmic domain of Notch contains nuclear localization sequences that function in cultured cells, and one of the Notch antineurogenic proteins, the cytoplasmic domain, accumulates in nuclei in vivo.
Lieber
T
T
Laboratory of Genetics, Rockefeller University, New York, New York 10021.
Kidd
S
S
Alcamo
E
E
Corbin
V
V
Young
M W
MW
D000818:Animals; D030801:Animals, Genetically Modified / Q000196:embryology; D002451:Cell Compartmentation; D018797:Cell Cycle Proteins; D002467:Cell Nucleus / Q000378:metabolism*; D003545:Cysteine / Q000235:genetics; D004252:DNA Mutational Analysis; D004330:Drosophila / Q000196:embryology* / Q000235:genetics; D029721:Drosophila Proteins; D004815:Epidermal Growth Factor / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics; D017344:Genes, Insect / Q000235:genetics; D007150:Immunohistochemistry; D007301:Insect Hormones / Q000096:biosynthesis / Q000235:genetics*; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics*; D009417:Nerve Tissue / Q000196:embryology*; D010446:Peptide Fragments / Q000302:isolation & purification; D010641:Phenotype; D051880:Receptors, Notch; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics; D015183:Restriction Mapping; D029702:Schizosaccharomyces pombe Proteins; D017384:Sequence Deletion; D015398:Signal Transduction; D014157:Transcription Factors / Q000235:genetics
D018797:Cell Cycle Proteins; D029721:Drosophila Proteins; D005656:Fungal Proteins; D007301:Insect Hormones; D008565:Membrane Proteins; C497759:N protein, Drosophila; D010446:Peptide Fragments; D051880:Receptors, Notch; D029702:Schizosaccharomyces pombe Proteins; D014157:Transcription Factors; C061178:cdc10 protein, S pombe; D004815:Epidermal Growth Factor; D003545:Cysteine