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首页> 美国卫生研究院文献>Frontiers in Plant Science >AtSIG6 and other members of the sigma gene family jointly but differentially determine plastid target gene expression in Arabidopsis thaliana
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AtSIG6 and other members of the sigma gene family jointly but differentially determine plastid target gene expression in Arabidopsis thaliana

机译：AtSIG6和sigma基因家族的其他成员共同但有区别地确定拟南芥中质体靶基因的表达

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摘要

Plants contain a nuclear gene family for plastid sigma factors, i.e., proteins that associate with the “bacterial-type” organellar RNA polymerase and confer the ability for correct promoter binding and transcription initiation. Questions that are still unresolved relate to the “division of labor” among members of the sigma family, both in terms of their range of target genes and their temporal and spatial activity during development. Clues to the in vivo role of individual sigma genes have mainly come from studies of sigma knockout lines. Despite its obvious strengths, however, this strategy does not necessarily trace-down causal relationships between mutant phenotype and a single sigma gene, if other family members act in a redundant and/or compensatory manner. We made efforts to reduce the complexity by genetic crosses of Arabidopsis single mutants (with focus on a chlorophyll-deficient sig6 line) to generate double knockout lines. The latter typically had a similar visible phenotype as the parental lines, but tended to be more strongly affected in the transcript patterns of both plastid and sigma genes. Because triple mutants were lethal under our growth conditions, we exploited a strategy of transformation of single and double mutants with RNAi constructs that contained sequences from the unconserved sigma region (UCR). These RNAi/knockout lines phenotypically resembled their parental lines, but were even more strongly affected in their plastid transcript patterns. Expression patterns of sigma genes revealed both similarities and differences compared to the parental lines, with transcripts at reduced or unchanged amounts and others that were found to be present in higher (perhaps compensatory) amounts. Together, our results reveal considerable flexibility of gene activity at the levels of both sigma and plastid gene expression. A (still viable) “basal state” seems to be reached, if 2–3 of the 6 Arabidopsis sigma genes are functionally compromised.
机译：植物包含质体sigma因子的核基因家族，即与“细菌型”细胞器RNA聚合酶相关的蛋白质，并赋予正确启动子结合和转录启动的能力。尚未解决的问题与sigma家族成员之间的“分工”有关，既涉及目标基因的范围，也涉及发育过程中的时空活动。各个sigma基因在体内作用的线索主要来自对sigma基因敲除系的研究。尽管具有明显的优势，但是，如果其他家族成员以多余和/或补偿的方式行事，则该策略不一定能追踪突变表型和单个sigma基因之间的因果关系。我们通过降低拟南芥单突变体（主要针对叶绿素缺陷型sig6品系）的遗传杂交以产生双敲除品系的方式来降低复杂性。后者通常具有与亲本系相似的可见表型，但是倾向于在质体和σ基因的转录模式中受到更强烈的影响。由于三重突变体在我们的生长条件下具有致死性，因此我们开发了一种利用RNAi构建体转化单突变体和双突变体的策略，该构建体包含来自非保守sigma区（UCR）的序列。这些RNAi /敲除系在表型上与它们的亲本系相似，但是在其质体转录物模式中甚至受到更大的影响。与亲本系相比，sigma基因的表达模式显示出相似性和差异性，其中转录本的数量减少或保持不变，而其他转录本的数量更高（也许是补偿性的）。总之，我们的结果显示出在sigma和质体基因表达水平上基因活性具有相当大的灵活性。如果6个拟南芥sigma基因中的2–3个在功能上受到损害，似乎已经达到（仍然可行的）“基础状态”。

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