Fitch C L, de Sousa S M, O'Day P M, Neubert T A, Plantilla C M, Spencer M, Yarfitz S, Apte D, Hurley J B
Howard Hughes Medical Institute, University of Washington, Seattle 98195.
Cell Signal. 1993 Mar;5(2):187-207. doi: 10.1016/0898-6568(93)90070-3.
Pertussis toxin inactivates certain G-proteins by introducing an ADP-ribose group near the carboxyl-terminus of the alpha-subunit. The major pertussis toxin substrate in Drosophila tissues is Go alpha. We introduced a pertussis toxin gene under control of the hsp70 heat-shock promoter into the Drosophila genome. When heat-shocked, transformed flies produce active pertussis toxin which ADP-ribosylates endogenous Go alpha. Pertussis toxin is expressed in photoreceptors, in the lamina of the eye and in epithelial cells lining the gut. As expected from the absence of Go alpha in photoreceptors, pertussis toxin does not affect the photoreceptor component of the Drosophila visual response. However, it abolishes light on- and off-transients in the electroretinogram. These transients normally arise from the lamina, a tissue where Go alpha transcripts have been detected. Pertussis toxin expression also blocks embryonic development and shortens the lifetime of adult Drosophila. Following heat-shock, transformed adults are active, but they fail to take up nutrients because they stop eating. High energy metabolites are significantly depleted shortly after pertussis toxin expression is induced and the flies die within 48 h.
百日咳毒素通过在α亚基羧基末端附近引入一个ADP - 核糖基团来使某些G蛋白失活。果蝇组织中主要的百日咳毒素底物是Goα。我们将一个受hsp70热休克启动子控制的百日咳毒素基因导入果蝇基因组。热休克时,转基因果蝇产生有活性的百日咳毒素,该毒素会对内源性Goα进行ADP - 核糖基化。百日咳毒素在光感受器、眼的神经纤维层以及肠道内衬上皮细胞中表达。正如光感受器中不存在Goα所预期的那样,百日咳毒素不影响果蝇视觉反应的光感受器成分。然而,它消除了视网膜电图中的光开和光关瞬变。这些瞬变通常起源于神经纤维层,在该组织中已检测到Goα转录本。百日咳毒素的表达还会阻断胚胎发育并缩短成年果蝇的寿命。热休克后,转基因成虫有活动能力,但它们因为停止进食而无法摄取营养。在诱导百日咳毒素表达后不久,高能代谢产物显著消耗,果蝇在48小时内死亡。