Tsolakis George, Moschonas Nicholas K, Galland Paul, Kotzabasis Kiriakos
Department of Biology, University of Crete, Crete, Herakliou, Greece.
Photochem Photobiol. 2004 Apr;79(4):360-70. doi: 10.1562/le-03-15.1.
Many responses of the zygomycete fungus Phycomyces blakesleeanus are mediated by blue light, e.g. the stimulation of beta-carotene synthesis (photocarotenogenesis) and the formation of fruiting bodies (photomorphogenesis). Even though both responses have been described in detail genetically and biophysically, the underlying molecular events remain unknown. Applying a pharmacological approach in developing mycelia, we investigated the possible involvement of heterotrimeric G proteins in the blue-light transduction chains of both responses. G protein agonists (guanosine triphosphate analogues, cholera toxin, pertussis toxin) mimicked in darkness the effect of blue light for both responses, except for cholera toxin, which was ineffective in increasing the beta-carotene content of dark-grown mycelia. Experiments combining the two toxins indicated that photocarotenogenesis could involve an inhibitory G protein (Gi) type, whereas photomorphogenesis may depend on a transducin (Gt type)-like heterotrimer. The determination of the carB (phytoene dehydrogenase) and chs1 (chitin synthase 1) gene expression under various conditions of exogenous challenge supports the G protein participation. The fluctuations of the time course measurements of the carB and chs1 transcripts are discussed.
接合菌纲真菌布氏毛霉的许多反应是由蓝光介导的,例如β-胡萝卜素合成的刺激(光致胡萝卜素生成)和子实体的形成(光形态发生)。尽管这两种反应在遗传学和生物物理学方面都已得到详细描述,但其潜在的分子事件仍然未知。在发育中的菌丝体中应用药理学方法,我们研究了异源三聚体G蛋白在这两种反应的蓝光转导链中可能的参与情况。G蛋白激动剂(鸟苷三磷酸类似物、霍乱毒素、百日咳毒素)在黑暗中模拟了蓝光对这两种反应的作用,但霍乱毒素除外,它对增加黑暗生长菌丝体的β-胡萝卜素含量无效。结合两种毒素的实验表明,光致胡萝卜素生成可能涉及一种抑制性G蛋白(Gi型),而光形态发生可能依赖于一种类似转导素(Gt型)的异源三聚体。在各种外源刺激条件下对carB(八氢番茄红素脱氢酶)和chs1(几丁质合酶1)基因表达的测定支持了G蛋白的参与。讨论了carB和chs1转录本的时间进程测量的波动情况。