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An AbrB-like protein might be involved in the regulation of

机译:AbrB样蛋白可能参与了

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Certain filamentous cyanobacteria, including Aphanizomenon ovalisporum, are potentially toxic owing to the formation of the hepatotoxin cylindrospermopsin. We previously identified a gene cluster in A. ovalisporum likely to be involved in cylindrospermopsin biosynthesis, including amidinotransferase (aoaA) and polyketide-synthase (aoaC), transcribed on the reverse strands. Analysis of the genomic region between aoaA and aoaC identified two transcription start points for each of these genes, differentially expressed under nitrogen and light stress conditions. The transcript abundances of these genes and the cylindrospermopsin level were both affected by nitrogen availability and light intensity. Gel shift assays and DNA affinity columns isolated a protein that specifically binds to a 150 bp DNA fragment from the region between aoaA and aoaC, and MS/MS analyses identified similarity to AbrB in other cyanobacteria and in Bacillus sp. Comparison of the native AbrB isolated from A. ovalisporum with that obtained after cloning and overexpression of abrB in Escherichia coli identified specific post-translational modifications in the native cyanobacterial protein. These modifications, which are missing in the protein expressed in E. coli, include N-acetylation and methylation of specific residues. We discuss the possible role of these modifications in the regulation of cylindrospermopsin production in Aphanizomenon.
机译:某些丝状蓝细菌,包括卵形隐孢单胞菌,由于形成了肝毒素的cylindrospermopsin而具有潜在的毒性。我们之前在卵形孢霉中发现了一个基因簇,可能与环精子藻生物素的生物合成有关,包括a链转移酶(aoaA)和聚酮化合物合酶(aoaC),转录在反向链上。对aoaA和aoaC之间的基因组区域的分析确定了这些基因中每个基因的两个转录起点,在氮和轻胁迫条件下差异表达。这些基因的转录丰度和圆柱精子蛋白水平都受到氮的有效性和光强度的影响。凝胶迁移分析和DNA亲和柱从aoaA和aoaC之间的区域中分离出一种与150 bp DNA片段特异性结合的蛋白质,MS / MS分析确定了与其他蓝细菌和芽孢杆菌属中AbrB的相似性。从椭圆形孢霉分离的天然AbrB与在大肠杆菌中克隆和过表达abrB后获得的天然AbrB的比较鉴定了天然蓝细菌蛋白的特定翻译后修饰。这些在大肠杆菌表达的蛋白质中缺失的修饰包括特定残基的N-乙酰化和甲基化。我们讨论了这些修饰在Aphanizomenon中对cysdrodrospermopsin生产的调节中的可能作用。

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