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首页> 外文期刊>Journal of bacteriology >A Putative O-Linked β-N-Acetylglucosamine Transferase Is Essential for Hormogonium Development and Motility in the Filamentous Cyanobacterium Nostoc punctiforme
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A Putative O-Linked β-N-Acetylglucosamine Transferase Is Essential for Hormogonium Development and Motility in the Filamentous Cyanobacterium Nostoc punctiforme

机译：推定的O-连接的β-N-乙酰氨基葡萄糖转移酶对于丝状蓝藻点状鼻孔菌的发育和运动至关重要。

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Most species of filamentous cyanobacteria are capable of gliding motility, likely via a conserved type IV pilus-like system that may also secrete a motility-associated polysaccharide. In a subset of these organisms, motility is achieved only after the transient differentiation of hormogonia, which are specialized filaments that enter a nongrowth state dedicated to motility. Despite the fundamental importance of hormogonia to the life cycles of many filamentous cyanobacteria, the molecular regulation of hormogonium development is largely undefined. To systematically identify genes essential for hormogonium development and motility in the model heterocyst-forming filamentous cyanobacterium Nostoc punctiforme, a forward genetic screen was employed. The first gene identified using this screen, designated ogtA, encodes a putative O-linked β-N-acetylglucosamine transferase (OGT). The deletion of ogtA abolished motility, while ectopic expression of ogtA induced hormogonium development even under hormogonium-repressing conditions. Transcription of ogtA is rapidly upregulated (1 h) following hormogonium induction, and an OgtA-GFPuv fusion protein localized to the cytoplasm. In developing hormogonia, accumulation of PilA but not HmpD is dependent on ogtA. Reverse transcription-quantitative PCR (RT-qPCR) analysis indicated equivalent levels of pilA transcript in the wild-type and ΔogtA mutant strains, while a reporter construct consisting of the intergenic region in the 5′ direction of pilA fused to gfp produced lower levels of fluorescence in the ΔogtA mutant strain than in the wild type. The production of hormogonium polysaccharide in the ΔogtA mutant strain is reduced compared to that in the wild type but comparable to that in a pilA deletion strain. Collectively, these results imply that O-GlcNAc protein modification regulates the accumulation of PilA via a posttranscriptional mechanism in developing hormogonia.
机译：大多数种类的丝状蓝细菌能够滑动运动，可能是通过保守的IV型菌毛样系统，该系统也可能分泌与运动相关的多糖。在这些生物的子集中，仅在短暂性的雄激素分化之后才能实现运动能力，而雄激素是进入运动专用的非生长状态的特殊长丝。尽管hormogonia对许多丝状蓝细菌的生命周期具有根本的重要性，但horgogonium发育的分子调控在很大程度上尚不确定。为了系统地识别模型中形成异囊的丝状蓝细菌点状诺氏菌中发育和运动所必需的基因，采用了正向遗传筛选。使用该筛选方法鉴定的第一个基因称为 ogtA ，编码一个假定的O-连接的β- N -乙酰氨基葡萄糖转移酶（OGT）。 ogtA 的缺失消除了运动力，而 ogtA 的异位表达即使在抑制mo的条件下也导致了mo的发育。在em诱导后， ogtA 的转录迅速上调（1小时），OgtA-GFPuv融合蛋白定位在细胞质中。在发生性激素血症中，PilA而不是HmpD的积累取决于 ogtA 。逆转录定量PCR（RT-qPCR）分析表明，野生型和Δ ogtA 突变株中 pilA 转录本的水平相同，而报告基因构建体由基因间与 gfp 融合的 pilA 5'方向的区域在Δ ogtA 突变菌株中产生的荧光水平低于野生型。与野生型相比，Δ ogtA 突变株中hor属多糖的产量减少，但与 pilA 缺失株中的mo属多糖的产量相比。总的来说，这些结果暗示 O -GlcNAc蛋白修饰可通过转录后机制调节发育过程中的PhorA积累。

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