Shalev-Malul Gali, Lieman-Hurwitz Judy, Viner-Mozzini Yehudith, Sukenik Assaf, Gaathon Ariel, Lebendiker Mario, Kaplan Aaron
Department of Plant and Environmental Sciences, The Hebrew University of Jerusalem, Jerusalem 91904, Israel.
Environ Microbiol. 2008 Apr;10(4):988-99. doi: 10.1111/j.1462-2920.2007.01519.x. Epub 2007 Dec 17.
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.
某些丝状蓝细菌,包括卵形隐球藻,由于会形成肝毒素柱孢藻毒素而具有潜在毒性。我们之前在卵形隐球藻中鉴定出一个可能参与柱孢藻毒素生物合成的基因簇,其中包括在反向链上转录的脒基转移酶(aoaA)和聚酮合酶(aoaC)。对aoaA和aoaC之间基因组区域的分析确定了这两个基因各自的两个转录起始点,它们在氮和光胁迫条件下差异表达。这些基因的转录丰度和柱孢藻毒素水平均受氮供应和光照强度的影响。凝胶迁移实验和DNA亲和柱分离出一种能特异性结合aoaA和aoaC之间区域150 bp DNA片段的蛋白质,质谱/质谱分析确定其与其他蓝细菌和芽孢杆菌属中的AbrB具有相似性。将从卵形隐球藻中分离出的天然AbrB与在大肠杆菌中克隆并过表达abrB后获得的AbrB进行比较,确定了天然蓝细菌蛋白中存在特定的翻译后修饰。这些在大肠杆菌中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卵形隐球藻中柱孢藻毒素产生调控中的可能作用。