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拟南芥中一种类似PII的蛋白质:在氮感知中的假定作用。

A PII-like protein in Arabidopsis: putative role in nitrogen sensing.

作者信息

Hsieh M H, Lam H M, van de Loo F J, Coruzzi G

机构信息

Department of Biology, New York University, New York, NY 10003, USA.

出版信息

Proc Natl Acad Sci U S A. 1998 Nov 10;95(23):13965-70. doi: 10.1073/pnas.95.23.13965.

Abstract

PII is a protein allosteric effector in Escherichia coli and other bacteria that indirectly regulates glutamine synthetase at the transcriptional and post-translational levels in response to nitrogen availability. Data supporting the notion that plants have a nitrogen regulatory system(s) includes previous studies showing that the levels of mRNA for plant nitrogen assimilatory genes such as glutamine synthetase (GLN) and asparagine synthetase (ASN) are modulated by carbon and organic nitrogen metabolites. Here, we have characterized a PII homolog (GLB1) in two higher plants, Arabidopsis thaliana and Ricinus communis (Castor bean). Each plant PII-like protein has high overall identity to E. coli PII (50%). Western blot analyses reveal that the plant PII-like protein is a nuclear-encoded chloroplast protein. The PII-like protein of plants appears to be regulated at the transcriptional level in that levels of GLB1 mRNA are affected by light and metabolites. To initiate studies of the in vivo function of the Arabidopsis PII-like protein, we have constructed transgenic lines in which PII expression is uncoupled from its native regulation. Analyses of these transgenic plants support the notion that the plant PII-like protein may serve as part of a complex signal transduction network involved in perceiving the status of carbon and organic nitrogen. Thus, the PII protein found in archaea, bacteria, and now in higher eukaryotes (plants) is one of the most widespread regulatory proteins known, providing evidence for an ancestral metabolic regulatory mechanism that may have existed before the divergence of these three domains of life.

摘要

PII是大肠杆菌和其他细菌中的一种蛋白质变构效应物,它根据氮的可利用性在转录和翻译后水平间接调节谷氨酰胺合成酶。支持植物具有氮调节系统这一观点的数据包括先前的研究,这些研究表明植物氮同化基因(如谷氨酰胺合成酶(GLN)和天冬酰胺合成酶(ASN))的mRNA水平受碳和有机氮代谢物的调节。在这里,我们鉴定了两种高等植物拟南芥和蓖麻(蓖麻子)中的PII同源物(GLB1)。每种植物类PII蛋白与大肠杆菌PII具有较高的整体同源性(50%)。蛋白质印迹分析表明,植物类PII蛋白是一种核编码的叶绿体蛋白。植物类PII蛋白似乎在转录水平受到调节,因为GLB1 mRNA的水平受光照和代谢物的影响。为了开始对拟南芥类PII蛋白的体内功能进行研究,我们构建了转基因系,其中PII的表达与其天然调节解偶联。对这些转基因植物的分析支持了这样一种观点,即植物类PII蛋白可能作为一个复杂信号转导网络的一部分,参与感知碳和有机氮的状态。因此,古细菌、细菌以及现在高等真核生物(植物)中发现的PII蛋白是已知分布最广泛的调节蛋白之一,这为可能在这三个生命域分化之前就已存在的祖先代谢调节机制提供了证据。

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