The Rac GTPase-activating protein RotundRacGAP interferes with Drac1 and Dcdc42 signalling in Drosophila melanogaster.
RhoGTPases are negatively regulated by GTPase-activating proteins (GAPs). Here we demonstrate that Drosophila RotundRacGAP is active in vitro on Drac1 and Dcdc42 but not Drho1. Similarly, in yeast, RotundRacGAP interacts specifically with Drac1 and Dcdc42, as well as with their activated V12 forms, showing a particularly strong interaction with Dcdc42V12. In the fly, lowering RotundRacGAP dosage specifically modifies eye defects induced by expressing Drac1 or Dcdc42 but not Drho1, confirming that Drac1 and Dcdc42 are indeed in vivo targets of RotundRacGAP. Furthermore, embryonic-directed expression of either RotundRacGAP, or dominant negative Drac1N17, transgenes induces similar defects in dorsal closure and inhibits Drac1-dependent cytoskeleton assembly at the leading edge. Expression of truncated forms of RotundRacGAP shows that the GAP domain of RotundRacGAP is essential for its function. Unexpectedly, transgenes encoding Drac1N17, Dcdc42N17, or RotundRacGAP do not affect the c-Jun N-terminal kinase-dependent gene expression of decapentaplegic and puckered, indicating that another Drac1-independent signal redundantly activates this pathway. Finally, in a situation where Drac1 is constitutively activated, RotundRacGAP greatly reduces the ectopic expression of decapentaplegic, possibly by negatively regulating Dcdc42.
Raymond
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Département de Biologie Moléculaire et Structurale, CEA-CNRS-UJF, UMR 5092, 17 rue des Martyrs, Grenoble 38054, France.
Bergeret
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Dagher
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MC
Breton
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Griffin-Shea
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Fauvarque
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MO
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D029721:Drosophila Proteins; D004331:Drosophila melanogaster; D004625:Embryo, Nonmammalian / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism; D020690:GTPase-Activating Proteins / Q000235:genetics / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D015398:Signal Transduction; D019076:Transgenes; D020744:rac GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*
C090145:Cdc42 protein, Drosophila; D017931:DNA Primers; D029721:Drosophila Proteins; D020690:GTPase-Activating Proteins; C090146:Rac1 protein, Drosophila; C434935:RacGAP84C protein, Drosophila; D019204:GTP-Binding Proteins; D020744:rac GTP-Binding Proteins