Swiatecka-Hagenbruch Monika, Emanuel Carola, Hedtke Boris, Liere Karsten, Börner Thomas
Institut für Biologie (Genetik), Humboldt-Universität zu Berlin, Chausseestrasse 117, D-10115 Berlin, Germany.
Nucleic Acids Res. 2008 Feb;36(3):785-92. doi: 10.1093/nar/gkm1111. Epub 2007 Dec 15.
Although chloroplast genomes are small, the transcriptional machinery is very complex in plastids of higher plants. Plastidial genes of higher plants are transcribed by plastid-encoded (PEP) and nuclear-encoded RNA polymerases (NEP). The nuclear genome of Arabidopsis contains two candidate genes for NEP, RpoTp and RpoTmp, both coding for phage-type RNA polymerases. We have analyzed the use of PEP and NEP promoters in transgenic Arabidopsis lines with altered RpoTp activities and in Arabidopsis RpoTp insertion mutants lacking functional RpoTp. Low or lacking RpoTp activity resulted in an albino phenotype of the seedlings, which normalized later in development. Differences in promoter usage between wild type and plants with altered RpoTp activity were also most obvious early in development. Nearly all NEP promoters were used in plants with low or lacking RpoTp activity, though certain promoters showed reduced or even increased usage. The strong NEP promoter of the essential ycf1 gene, however, was not used in mutant seedlings lacking RpoTp activity. Our data provide evidence for NEP being represented by two phage-type RNA polymerases (RpoTp and RpoTmp) that have overlapping as well as gene-specific functions in the transcription of plastidial genes.
尽管叶绿体基因组较小,但高等植物质体中的转录机制非常复杂。高等植物的质体基因由质体编码的(PEP)和核编码的RNA聚合酶(NEP)转录。拟南芥的核基因组包含两个NEP候选基因,RpoTp和RpoTmp,两者都编码噬菌体类型的RNA聚合酶。我们分析了RpoTp活性改变的转基因拟南芥品系以及缺乏功能性RpoTp的拟南芥RpoTp插入突变体中PEP和NEP启动子的使用情况。RpoTp活性低或缺乏导致幼苗白化表型,该表型在发育后期恢复正常。野生型与RpoTp活性改变的植株之间启动子使用的差异在发育早期也最为明显。几乎所有NEP启动子都在RpoTp活性低或缺乏的植株中使用,尽管某些启动子的使用有所减少甚至增加。然而,必需的ycf1基因的强NEP启动子在缺乏RpoTp活性的突变幼苗中未被使用。我们的数据为NEP由两种噬菌体类型的RNA聚合酶(RpoTp和RpoTmp)代表提供了证据,它们在质体基因转录中具有重叠以及基因特异性功能。