Foster K W, Saranak J, Zarrilli G
Physics Department, Syracuse University, NY 13244-1130.
Proc Natl Acad Sci U S A. 1988 Sep;85(17):6379-83. doi: 10.1073/pnas.85.17.6379.
A sensitive assay for the induction of carotenoid and rhodopsin synthesis, based on the phototactic response, has been developed in a mutant of the unicellular alga Chlamydomonas reinhardtii. In the dark, the mutant fails to synthesize carotene and retinal, but it contains the apoprotein opsin. When retinal synthesis is induced by light treatment, the retinal combines with opsin to form rhodopsin, and the cells swim away from a source of light. Since the amount of light required to trigger a phototactic response is inversely proportional to the concentration of rhodopsin, the decrease in amount of light necessary to generate that response can serve as a measure of the amount of retinal synthesized in cells after induction. Using this assay, we found that (i) light induction of retinal depends linearly on light exposure and rhodopsin concentration during the exposure; (ii) the action spectrum of light induction is identical with that for phototaxis for which the receptor pigment is rhodopsin; and (iii) incubation with all-trans-7,8-dihydroretinal before light exposure shifts the action-spectrum peak for light induction 0.41 eV (-71 nm). We conclude that the photopigment for induction of retinal synthesis is a rhodopsin. The time lag required for induction of retinal synthesis and preliminary experiments with transcription or translation inhibitors suggest that alterations in gene expression could be involved in the induction process. Its control could be similar to other processes in which membrane receptors for hormones, neurotransmitters, or growth factors regulate gene expression.
基于趋光反应,已在单细胞绿藻莱茵衣藻的一个突变体中开发出一种用于诱导类胡萝卜素和视紫红质合成的灵敏测定法。在黑暗中,该突变体无法合成胡萝卜素和视黄醛,但它含有脱辅基蛋白视蛋白。当通过光照处理诱导视黄醛合成时,视黄醛与视蛋白结合形成视紫红质,细胞会游向远离光源的方向。由于引发趋光反应所需的光量与视紫红质的浓度成反比,因此产生该反应所需光量的减少可作为诱导后细胞中合成的视黄醛量的一种度量。使用这种测定法,我们发现:(i)视黄醛的光诱导与光照期间的光照暴露和视紫红质浓度呈线性关系;(ii)光诱导的作用光谱与以视紫红质为受体色素的趋光性的作用光谱相同;(iii)在光照暴露前用全反式-7,8-二氢视黄醛孵育会使光诱导的作用光谱峰移动0.41电子伏特(-71纳米)。我们得出结论,诱导视黄醛合成的光色素是一种视紫红质。视黄醛合成诱导所需的时间滞后以及用转录或翻译抑制剂进行的初步实验表明,基因表达的改变可能参与了诱导过程。其调控可能类似于激素、神经递质或生长因子的膜受体调节基因表达的其他过程。