Ishii Ai, Hihara Yukako
Department of Biochemistry and Molecular Biology, Graduate School of Science and Engineering, Saitama University, Sakura-ku, Saitama 338-8570, Japan.
Plant Physiol. 2008 Sep;148(1):660-70. doi: 10.1104/pp.108.123505. Epub 2008 Jul 30.
Every cyanobacterial species possesses multiple genes encoding AbrB-like transcriptional regulators (cyAbrBs) distinct from those conserved among other bacterial species. In this study, two genes encoding cyAbrBs in Synechocystis sp. PCC 6803, sll0359 and sll0822, were insertionally disrupted in order to examine their physiological roles. A fully segregated disrupted mutant of sll0822 (Deltasll0822 mutant) but not of sll0359 was obtained, although both mutants exhibited similar phenotypes (i.e. decreases in growth rate and pigment content). The growth rate of the Deltasll0822 mutant was low under any condition, but the low pigment content could be partially recovered by nitrate supplementation of the medium. DNA microarray and RNA-blot analyses revealed that the level of expression of a part of the NtcA regulon, such as urtA, amt1, glnB, sigE, and the nrt operon, was significantly decreased in the Deltasll0822 mutant, although the induction of these genes upon nitrogen depletion was still observed to some extent. Sll0822 seems to work in parallel with NtcA to achieve flexible regulation of the nitrogen uptake system. The Sll0822 protein exists mainly in a dimeric form in vivo, and the amount of the protein was not affected by nitrogen availability. This observation, together with the low binding specificity of the purified histidine-tagged Sll0822 protein, implies that the activity of Sll0822 may be posttranslationally modulated in Synechocystis cells.
每种蓝藻物种都拥有多个编码AbrB样转录调节因子(cyAbrBs)的基因,这些基因与其他细菌物种中保守的基因不同。在本研究中,为了研究集胞藻PCC 6803中编码cyAbrBs的两个基因sll0359和sll0822的生理作用,对其进行了插入失活。尽管两个突变体表现出相似的表型(即生长速率和色素含量降低),但获得了sll0822的完全分离的失活突变体(Deltasll0822突变体),而sll0359的突变体未获得。Deltasll0822突变体在任何条件下生长速率都很低,但通过向培养基中添加硝酸盐,色素含量的降低可部分恢复。DNA微阵列和RNA印迹分析表明,在Deltasll0822突变体中,NtcA调控子的一部分基因(如urtA、amt1、glnB、sigE和nrt操纵子)的表达水平显著降低,尽管在一定程度上仍观察到这些基因在氮耗尽时的诱导。Sll0822似乎与NtcA协同作用,以实现对氮吸收系统的灵活调节。Sll0822蛋白在体内主要以二聚体形式存在,其含量不受氮供应情况的影响。这一观察结果,连同纯化的组氨酸标签Sll0822蛋白的低结合特异性,表明Sll0822的活性可能在集胞藻细胞中受到翻译后调控。