Zheng Chong-Ke, Wang Chun-Lian, Zhang Xiao-Ping, Wang Fu-Jun, Qin Teng-Fei, Zhao Kai-Jun
National Key Facility for Crop Gene Resources and Genetic Improvement (NFCRI), Key Laboratory of Crop Genetics and Breeding, Ministry of Agriculture, Institute of Crop Science, Chinese Academy of Agricultural Sciences (CAAS), Beijing, 100081, China.
Mol Plant Pathol. 2014 Sep;15(7):690-7. doi: 10.1111/mpp.12125. Epub 2014 Apr 10.
To activate the expression of host genes that contribute to pathogen growth, pathogenic Xanthomonas bacteria inject their transcription activator-like effectors (TALEs) into plant cells and the TALEs bind to target gene promoters by the central repeat region consisting of near-perfect 34-amino-acid repeats (34-aa repeats). Based on the recognition codes between the 34-aa repeats and the targeted nucleotides, TALE-based technologies, such as designer TALEs (dTALEs) and TALE nucleases (TALENs), have been developed. Amazingly, every natural TALE invariantly has a truncated last half-repeat (LHR) at the end of the 34-aa repeats. Consequently, all the reported dTALEs and TALENs also harbour their LHRs. Here, we show that the LHRs in dTALEs are dispensable for the function of gene activation by both transient expression assays in Nicotiana benthamiana and gene-specific targeting in the rice genome, indicating that TALEs might originate from a single progenitor. In the light of this finding, we demonstrate that dTALEs can be constructed through two simple steps. Moreover, the activation strengths of dTALEs lacking the LHR are comparable with those of dTALEs harbouring the LHR. Our results provide new insights into the origin of natural TALEs, and will facilitate the simplification of the design and assembly of TALE-based tools, such as dTALEs and TALENs, in the near future.
为激活有助于病原体生长的宿主基因的表达,致病性黄单胞菌将其转录激活样效应因子(TALEs)注入植物细胞,这些TALEs通过由近乎完美的34个氨基酸重复序列(34-aa重复序列)组成的中央重复区域与靶基因启动子结合。基于34-aa重复序列与靶向核苷酸之间的识别密码,已开发出基于TALE的技术,如定制TALEs(dTALEs)和TALE核酸酶(TALENs)。令人惊讶的是,每个天然TALE在34-aa重复序列末端都始终有一个截短的后半重复序列(LHR)。因此,所有报道的dTALEs和TALENs也都含有它们的LHR。在这里,我们通过在本氏烟草中的瞬时表达分析以及在水稻基因组中的基因特异性靶向研究表明,dTALEs中的LHR对于基因激活功能是可有可无的,这表明TALEs可能起源于单个祖先。鉴于这一发现,我们证明可以通过两个简单步骤构建dTALEs。此外,缺乏LHR的dTALEs的激活强度与含有LHR的dTALEs相当。我们的结果为天然TALEs的起源提供了新的见解,并将在不久的将来促进基于TALE的工具(如dTALEs和TALENs)的设计和组装的简化。