Mussgnug Jan H, Wobbe Lutz, Elles Ingolf, Claus Christina, Hamilton Mary, Fink Andreas, Kahmann Uwe, Kapazoglou Aliki, Mullineaux Conrad W, Hippler Michael, Nickelsen Jörg, Nixon Peter J, Kruse Olaf
Department of Biology/Molecular Cell Physiology, University of Bielefeld, Germany.
Plant Cell. 2005 Dec;17(12):3409-21. doi: 10.1105/tpc.105.035774. Epub 2005 Nov 11.
Photosynthetic organisms respond to changes in ambient light by modulating the size and composition of their light-harvesting complexes, which in the case of the green alga Chlamydomonas reinhardtii consists of >15 members of a large extended family of chlorophyll binding subunits. How their expression is coordinated is unclear. Here, we describe the analysis of an insertion mutant, state transitions mutant3 (stm3), which we show has increased levels of LHCBM subunits associated with the light-harvesting antenna of photosystem II. The mutated nuclear gene in stm3 encodes the RNA binding protein NAB1 (for putative nucleic acid binding protein). In vitro and in vivo RNA binding and protein expression studies have confirmed that NAB1 differentially binds to LHCBM mRNA in a subpolysomal high molecular weight RNA-protein complex. Binding of NAB1 stabilizes LHCBM mRNA at the preinitiation level via sequestration and thereby represses translation. The specificity and affinity of binding are determined by an RNA sequence motif similar to that used by the Xenopus laevis translation repressor FRGY2, which is conserved to varying degrees in the LHCBM gene family. We conclude from our results that NAB1 plays an important role in controlling the expression of the light-harvesting antenna of photosystem II at the posttranscriptional level. The similarity of NAB1 and FRGY2 of Xenopus implies the existence of similar RNA-masking systems in animals and plants.
光合生物通过调节其捕光复合体的大小和组成来响应环境光的变化,对于绿藻莱茵衣藻来说,其捕光复合体由一个大型扩展叶绿素结合亚基家族的15个以上成员组成。目前尚不清楚它们的表达是如何协调的。在此,我们描述了对一个插入突变体——状态转换突变体3(stm3)的分析,我们发现该突变体中与光系统II捕光天线相关的LHCBM亚基水平有所增加。stm3中发生突变的核基因编码RNA结合蛋白NAB1(假定的核酸结合蛋白)。体外和体内的RNA结合及蛋白质表达研究证实,NAB1在一个亚多聚体高分子量RNA-蛋白质复合物中与LHCBM mRNA存在差异结合。NAB1的结合通过隔离作用在起始前水平稳定LHCBM mRNA,从而抑制翻译。结合的特异性和亲和力由一个与非洲爪蟾翻译抑制因子FRGY2所使用的RNA序列基序相似的序列决定,该基序在LHCBM基因家族中存在不同程度的保守性。我们从结果中得出结论,NAB1在转录后水平控制光系统II捕光天线的表达中起重要作用。NAB1与非洲爪蟾FRGY2的相似性意味着动植物中存在类似的RNA掩盖系统。