Functional Analysis of the Channelrhodopsin Genes from the Green Algae of the White Sea Basin

IF 2.3 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Olga V. Karpova, Elizaveta N. Vinogradova, Anastasiya M. Moisenovich, Oksana B. Pustovit, Alla A. Ramonova, Denis V. Abramochkin, Elena S. Lobakova
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Abstract

Optogenetics, the method of light-controlled regulation of cellular processes is based on the use of the channelrhodopsins that directly generate photoinduced currents. Most of the channelrhodopsin genes have been identified in the green microalgae Chlorophyta, and the demand for increasing the number of functionally characterized channelrhodopsins and the diversity of their photochemical parameters keeps growing. We performed the expression analysis of cation channelrhodopsin (CCR) genes in natural isolates of microalgae of the genera Haematococcus and Bracteacoccus from the unique Arctic Circle region. The identified full-length CCR transcript of H. lacustris is the product of alternative splicing and encodes the Hl98CCR2 protein with no photochemical activity. The 5′-partial fragment of the B. aggregatus CCR transcript encodes the Ba34CCR protein containing a conserved TM1-TM7 membrane domain and a short cytosolic fragment. Upon heterologous expression of the TM1-TM7 fragment in CHO-K1 cell culture, light-dependent current generation was observed with the parameters corresponding to those of the CCR. The first discovered functional channelrhodopsin of Bracteacoccus has no close CCR homologues and may be of interest as a candidate for optogenetics.

Abstract Image

白海盆地绿藻中的通道荧光素基因的功能分析。
光遗传学(Optogenetics)是一种通过光控调节细胞过程的方法,其基础是利用直接产生光诱导电流的通道闪烁蛋白。目前已在绿色微藻类叶绿藻中发现了大部分通道闪烁蛋白基因,而且对增加具有功能特征的通道闪烁蛋白数量及其光化学参数多样性的需求也在不断增长。我们对来自北极圈地区的血球藻属(Haematococcus)和白桦球藻属(Bracteacoccus)微藻天然分离物中的阳离子通道荧光素(CCR)基因进行了表达分析。已确定的 H. lacustris 的全长 CCR 转录本是替代剪接的产物,编码没有光化学活性的 Hl98CCR2 蛋白。B. aggregatus CCR 转录本的 5'-partial 片段编码 Ba34CCR 蛋白,其中包含一个保守的 TM1-TM7 膜结构域和一个短的细胞膜片段。在 CHO-K1 细胞培养中异源表达 TM1-TM7 片段后,观察到了光依赖性电流产生,其参数与 CCR 的参数一致。首次发现的白racteacoccus 功能性通道视蛋白没有近似的 CCR 同源物,可能是光遗传学的候选物质。
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来源期刊
Biochemistry (Moscow)
Biochemistry (Moscow) 生物-生化与分子生物学
CiteScore
4.70
自引率
3.60%
发文量
139
审稿时长
2 months
期刊介绍: Biochemistry (Moscow) is the journal that includes research papers in all fields of biochemistry as well as biochemical aspects of molecular biology, bioorganic chemistry, microbiology, immunology, physiology, and biomedical sciences. Coverage also extends to new experimental methods in biochemistry, theoretical contributions of biochemical importance, reviews of contemporary biochemical topics, and mini-reviews (News in Biochemistry).
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