RESUMEN
During embryogenesis, the activated Torso receptor tyrosine kinase (TOR RTK) pathway activates tailless (tll) expression by a relief-of-repression mechanism. Various lines of evidence have suggested that multiple factors are required for this repression. We show that Tramtrack69 (TTK69) binds to two sites within tll cis-regulatory DNA, TC2 and TC5, and that TTK69 is phosphorylated by mitogen activated protein kinase. In embryos lacking maternal ttk69 activity, the expression of both endogenous tll and lacZ driven by the tll minimal regulatory region (tll-MRR) are expanded. Further, in wild-type embryos, the tll-MRR mutated in TC5 drives uniform lacZ expression before late stage 4. We conclude that TTK69 is required for early (before the end of stage 4) repression of tll transcription.
Asunto(s)
Proteínas de Unión al ADN/genética , Proteínas de Drosophila/genética , Drosophila/embriología , Drosophila/genética , Proteínas Represoras/genética , Proteínas Represoras/metabolismo , Animales , Animales Modificados Genéticamente , Secuencia de Bases , Sitios de Unión/genética , ADN/genética , ADN/metabolismo , Drosophila/metabolismo , Proteínas de Drosophila/metabolismo , Femenino , Regulación del Desarrollo de la Expresión Génica , Genes de Insecto , Operón Lac , Masculino , Mutación , Proteínas Tirosina Quinasas Receptoras/metabolismoRESUMEN
Multidrug resistance (MDR) genes are abundant in Streptomyces genomes, and yet these bacteria are generally drug sensitive under routine laboratory conditions, indicating low or no expression of these genes. Drug-resistant mutations have been isolated that lie in regulatory genes adjacent to the MDR genes, suggesting that resistance arises by derepression. This study identified a divergently oriented pair consisting of a TetR-family regulator (ebrS) and a major facilitator-family MDR pump (ebrC) gene in Streptomyces lividans, which is widely conserved in Streptomyces species. EbrS represses transcription of ebrC as well as its own transcription. Deletion of ebrS causes overexpression of ebrC, resulting in elevated resistance to many drugs. The ebrS and ebrC promoters were used in a reporter system to test inducibility by various chemicals. Among the 15 compounds (including five EbrC target drugs) tested, none induced ebrC transcription. On the other hand, the ebrS promoter was induced by rifampicin and high concentrations of calcium and magnesium. Deletion of ebrS-ebrC did not change rifampicin sensitivity, indicating that the EbrC pump is not involved in rifampicin efflux. Moreover, deletion of ebrC caused retardation of colony growth on selected media, and the defect could be suppressed by supplementation with high concentrations of Ca(2+), Mg(2+), Na(+) or K(+). Based on these results, it is proposed that the primary biological role of most MDR systems in Streptomyces species is not removal of extrinsic drugs, but rather export of specific toxic compounds endogenously synthesized during growth.