Ebnet E, Fischer M, Deininger W, Hegemann P
Institut für Biochemie I, Universität Regensburg, 93040 Regensburg, Germany.
Plant Cell. 1999 Aug;11(8):1473-84. doi: 10.1105/tpc.11.8.1473.
Somatic cells of the multicellular alga Volvox carteri contain a visual rhodopsin that controls the organism's phototactic behavior via two independent photoreceptor currents. Here, we report the identification of an opsinlike gene, designated as volvoxopsin (vop). The encoded protein exhibits homologies to the opsin of the unicellular alga Chlamydomonas reinhardtii (chlamyopsin) and to the entire animal opsin family, thus providing new perspectives on opsin evolution. Volvoxopsin accumulates within the eyes of somatic cells. However, the vop transcript is detectable only in the reproductive eyeless gonidia and embryos. vop mRNA levels increase 400-fold during embryogenesis, when embryos develop in darkness, whereas the vop transcript does not accumulate when embryos develop in the light. An antisense transformant, T3, was generated. This transformant produces 10 times less volvoxopsin than does the wild type. In T3, the vop transcript is virtually absent, whereas the antisense transcript is predominant and light regulated. It follows that vop expression is under light-dependent transcriptional control but that volvoxopsin itself is not the regulatory photoreceptor. Transformant T3 is phototactic, but its phototactic sensitivity is reduced 10-fold relative to the parental wild-type strain HK10. Thus, we offer definitive genetic evidence that a rhodopsin serves as the photoreceptor for phototaxis in a green alga.
多细胞藻类团藻的体细胞含有一种视觉视紫红质,它通过两种独立的光感受器电流来控制生物体的趋光行为。在此,我们报告了一个类视蛋白基因的鉴定,该基因被命名为团藻视蛋白(vop)。编码的蛋白质与单细胞藻类莱茵衣藻的视蛋白(衣藻视蛋白)以及整个动物视蛋白家族具有同源性,从而为视蛋白的进化提供了新的视角。团藻视蛋白在体细胞的眼点内积累。然而,vop转录本仅在无眼的生殖性生殖细胞和胚胎中可检测到。在胚胎发育过程中,当胚胎在黑暗中发育时,vop mRNA水平增加400倍,而当胚胎在光照下发育时,vop转录本不会积累。构建了一个反义转化体T3。该转化体产生的团藻视蛋白比野生型少10倍。在T3中,几乎不存在vop转录本,而反义转录本占主导且受光调节。由此可见,vop的表达受光依赖的转录控制,但团藻视蛋白本身并不是调节性光感受器。转化体T3具有趋光性,但其趋光敏感性相对于亲本野生型菌株HK10降低了10倍。因此,我们提供了确凿的遗传学证据,证明视紫红质在绿藻中作为趋光性的光感受器。