Relationships between protein isoforms and genetic functions demonstrate functional redundancy at the Broad-Complex during Drosophila metamorphosis.
Metamorphosis in holometabolous insects is an ecdysone-dependent process by which the larval form is replaced by a reproductive, adult form. At the onset of metamorphosis ecdysone induces a set of early genes which coordinate tissue-specific responses to hormone. The Broad-Complex (BR-C) early gene, which acts as a global regulator of tissue-specific responses to ecdysone, encodes a family of zinc-finger DNA binding proteins known as Z1, Z2, Z3, and Z4. Genetically the BR-C encodes three complementing functions, br, rbp, and 2Bc, and a class of npr1 alleles that fail to complement any of the other genetic functions. The effects of BR-C mutations on metamorphic development are highly pleiotropic, yet little is known about the roles of individual BR-C proteins in directing the required responses to ecdysone. Because the BR-C is a vital regulator of metamorphosis it is essential to establish the relationships between BR-C genetic functions and protein products. We present here the first general and definitive study of these relationships. Using heat-inducible transgenes we have rescued lethality associated with each of the complementing genetic functions and have restored transcriptional activity of tissue-specific BR-C(+)-dependent target genes. Our data lead us to conclude that br+ function is only provided by the Z2 isoform. We find that Z1 transgenes provide full rbp+ function, while Z4 provides partial function. Likewise, while Z3 provides full 2Bc+ function, Z2 also provides partial function. These results indicate possible functional redundancy or regulatory dependence (via autoregulation) associated with the rbp+ and 2Bc+ functions. The establishment of these relationships between BR-C genetic functions and protein isoforms is an important step toward understanding the roles of BR-C proteins in directing metamorphic responses to ecdysone.
Bayer
C A
CA
Department of Molecular and Cell Biology, University of California, Berkeley 94720-3200, USA. cbayer@pegasus.cc.ucf.edu
von Kalm
L
L
Fristrom
J W
JW
D000426:Alcohol Dehydrogenase / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D030801:Animals, Genetically Modified; D002490:Central Nervous System / Q000378:metabolism; D004296:Dopa Decarboxylase / Q000096:biosynthesis / Q000235:genetics; D004330:Drosophila / Q000196:embryology / Q000235:genetics* / Q000254:growth & development; D029721:Drosophila Proteins; D004440:Ecdysone / Q000378:metabolism; D005216:Fat Body / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D017344:Genes, Insect; D005804:Genes, Lethal; D005816:Genetic Complementation Test; D005967:Glue Proteins, Drosophila / Q000096:biosynthesis / Q000235:genetics; D008297:Male; D008675:Metamorphosis, Biological / Q000235:genetics*; D009154:Mutation; D012469:Salivary Glands / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics*; D016335:Zinc Fingers / Q000235:genetics*
C102117:Br protein, Drosophila; D029721:Drosophila Proteins; D005967:Glue Proteins, Drosophila; D014157:Transcription Factors; D004440:Ecdysone; D000426:Alcohol Dehydrogenase; D004296:Dopa Decarboxylase