Kunihiro Shuta, Kowata Hikaru, Kondou Youichi, Takahashi Shinya, Matsui Minami, Berberich Thomas, Youssefian Shohab, Hidema Jun, Kusano Tomonobu
Graduate School of Life Sciences, Tohoku University, 2-1-1 Katahira, Aoba, Sendai, Miyagi, 980-8577, Japan.
Planta. 2014 May;239(5):1101-11. doi: 10.1007/s00425-014-2042-1. Epub 2014 Feb 23.
Screening of 40,000 Arabidopsis FOX (Full-length cDNA Over-eXpressor gene hunting system) lines expressing rice full-length cDNAs brings us to identify four cadmium (Cd)-tolerant lines, one of which carried OsREX1-S as a transgene. OsREX1-S shows the highest levels of identity to Chlamydomonas reinhardtii REX1-S (referred to as CrREX1-S, in which REX denotes Required for Excision) and to yeast and human TFB5s (RNA polymerase II transcription factor B5), both of which are components of the general transcription and DNA repair factor, TFIIH. Transient expression of OsREX1-S consistently localized the protein to the nucleus of onion cells. The newly generated transgenic Arabidopsis plants expressing OsREX1-S reproducibly displayed enhanced Cd tolerance, confirming that the Cd-tolerance of the initial identified line was conferred solely by OsREX1-S expression. Furthermore, transgenic Arabidopsis plants expressing OsREX1-S exhibited ultraviolet-B (UVB) tolerance by reducing the amounts of cyclobutane pyrimidine dimers produced by UVB radiation. Moreover, those transgenic OsREX1-S Arabidopsis plants became resistant to bleomycin (an inducer of DNA strand break) and mitomycin C (DNA intercalating activity), compared to wild type. Our results indicate that OsREX1-S renders host plants tolerant to Cd, UVB radiation, bleomycin and mitomycin C through the enhanced DNA excision repair.
对40000个表达水稻全长cDNA的拟南芥FOX(全长cDNA过表达基因筛选系统)株系进行筛选,使我们鉴定出4个耐镉株系,其中一个携带OsREX1-S作为转基因。OsREX1-S与莱茵衣藻REX1-S(称为CrREX1-S,其中REX表示切除所需)以及酵母和人类TFB5s(RNA聚合酶II转录因子B5)具有最高的同源性,这两者都是一般转录和DNA修复因子TFIIH的组成部分。OsREX1-S的瞬时表达始终将该蛋白定位在洋葱细胞的细胞核中。新产生的表达OsREX1-S的转基因拟南芥植株可重复地表现出增强的耐镉性,证实最初鉴定的株系的耐镉性仅由OsREX1-S的表达赋予。此外,表达OsREX1-S的转基因拟南芥植株通过减少UVB辐射产生的环丁烷嘧啶二聚体的量而表现出对紫外线B(UVB)的耐受性。此外,与野生型相比,那些转基因OsREX1-S拟南芥植株对博来霉素(一种DNA链断裂诱导剂)和丝裂霉素C(DNA嵌入活性)具有抗性。我们的结果表明,OsREX1-S通过增强DNA切除修复使宿主植物对镉、UVB辐射、博来霉素和丝裂霉素C具有耐受性。