Visualization of Nuclear Localization of Transcription Factors with Cyan and Green Fluorescent Proteins in the Red Alga Porphyra yezoensis
Visualization of Nuclear Localization of Transcription Factors with Cyan and Green Fluorescent Proteins in the Red Alga Porphyra yezoensis
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DOI：
10.1007/s10126-009-9210-5
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发表时间：
2010-04-01
影响因子：
3
通讯作者：
Mikami, Koji
中科院分区：
文献类型：
--
作者：
Uji, Toshiki;Takahashi, Megumu;Mikami, Koji
转录因子在所有生物体中基因组信息的表达中起核心作用。本研究的目的是分析红藻中转录因子的功能。分析真核细胞中转录因子的一种方法是研究它们的核定位，如使用荧光蛋白对陆地植物和绿色藻类的报道。然而，目前还没有报道记录红藻转录因子的亚细胞定位。在本研究中，我们使用海洋红藻条斑紫菜，首次证实了来自珊瑚Zoanthus sp.的人源化荧光蛋白（ZsGFP和ZsYFP）和陆地植物适应性sGFP（S65 T）在配子体细胞中的成功表达，与AmCFP的表达相当。在转录因子DP-E2 F-like 1（PyDEL 1）、转录延伸因子1（PyElf 1）和多蛋白桥接因子1（PyMBF 1）的分子克隆和表征之后，我们证明了ZsGFP和AmCFP可以用于可视化PyElf 1和PyMBF 1的核定位。这是第一份报告，以执行可视化的亚细胞定位的转录因子作为基因组编码的蛋白质在红藻。
Transcription factors play a central role in expression of genomic information in all organisms. The objective of our study is to analyze the function of transcription factors in red algae. One way to analyze transcription factors in eukaryotic cells is to study their nuclear localization, as reported for land plants and green algae using fluorescent proteins. There is, however, no report documenting subcellular localization of transcription factors from red algae. In the present study, using the marine red alga Porphyra yezoensis, we confirmed for the first time successful expression of humanized fluorescent proteins (ZsGFP and ZsYFP) from a reef coral Zoanthus sp. and land plant-adapted sGFP(S65T) in gametophytic cells comparable to expression of AmCFP. Following molecular cloning and characterization of transcription factors DP-E2F-like 1 (PyDEL1), transcription elongation factor 1 (PyElf1) and multiprotein bridging factor 1 (PyMBF1), we then demonstrated that ZsGFP and AmCFP can be used to visualize nuclear localization of PyElf1 and PyMBF1. This is the first report to perform visualization of subcellular localization of transcription factors as genome-encoded proteins in red algae.