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特定的葡萄糖苷转运蛋白影响丝状、形成异形胞的蓝藻鱼腥藻PCC 7120菌株的中隔结构和功能。

Specific Glucoside Transporters Influence Septal Structure and Function in the Filamentous, Heterocyst-Forming Cyanobacterium Anabaena sp. Strain PCC 7120.

作者信息

Nieves-Morión Mercedes, Lechno-Yossef Sigal, López-Igual Rocío, Frías José E, Mariscal Vicente, Nürnberg Dennis J, Mullineaux Conrad W, Wolk C Peter, Flores Enrique

机构信息

Instituto de Bioquímica Vegetal y Fotosíntesis, CSIC and Universidad de Sevilla, Seville, Spain.

MSU-DOE Plant Research Laboratory, Michigan State University, East Lansing, Michigan, USA.

出版信息

J Bacteriol. 2017 Mar 14;199(7). doi: 10.1128/JB.00876-16. Print 2017 Apr 1.

Abstract

When deprived of combined nitrogen, some filamentous cyanobacteria contain two cell types: vegetative cells that fix CO through oxygenic photosynthesis and heterocysts that are specialized in N fixation. In the diazotrophic filament, the vegetative cells provide the heterocysts with reduced carbon (mainly in the form of sucrose) and heterocysts provide the vegetative cells with combined nitrogen. Septal junctions traverse peptidoglycan through structures known as nanopores and appear to mediate intercellular molecular transfer that can be traced with fluorescent markers, including the sucrose analog esculin (a coumarin glucoside) that is incorporated into the cells. Uptake of esculin by the model heterocyst-forming cyanobacterium sp. strain PCC 7120 was inhibited by the α-glucosides sucrose and maltose. Analysis of mutants identified components of three glucoside transporters that move esculin into the cells: GlsC (Alr4781) and GlsP (All0261) are an ATP-binding subunit and a permease subunit of two different ABC transporters, respectively, and HepP (All1711) is a major facilitator superfamily (MFS) protein that was shown previously to be involved in formation of the heterocyst envelope. Transfer of fluorescent markers (especially calcein) between vegetative cells of was impaired by mutation of glucoside transporter genes. GlsP and HepP interact in bacterial two-hybrid assays with the septal junction-related protein SepJ, and GlsC was found to be necessary for the formation of a normal number of septal peptidoglycan nanopores and for normal subcellular localization of SepJ. Therefore, beyond their possible role in nutrient uptake in , glucoside transporters influence the structure and function of septal junctions. Heterocyst-forming cyanobacteria have the ability to perform oxygenic photosynthesis and to assimilate atmospheric CO and N These organisms grow as filaments that fix these gases specifically in vegetative cells and heterocysts, respectively. For the filaments to grow, these types of cells exchange nutrients, including sucrose, which serves as a source of reducing power and of carbon skeletons for the heterocysts. Movement of sucrose between cells in the filament takes place through septal junctions and has been traced with a fluorescent sucrose analog, esculin, that can be taken up by the cells. Here, we identified α-glucoside transporters of that mediate uptake of esculin and, notably, influence septal structure and the function of septal junctions.

摘要

当缺乏化合态氮时,一些丝状蓝细菌含有两种细胞类型:通过有氧光合作用固定二氧化碳的营养细胞和专门进行固氮的异形胞。在固氮丝状体中,营养细胞为异形胞提供还原态碳(主要以蔗糖形式),而异形胞为营养细胞提供化合态氮。隔膜连接通过称为纳米孔的结构穿过肽聚糖,似乎介导细胞间分子转移,这种转移可用荧光标记物追踪,包括掺入细胞中的蔗糖类似物七叶苷(一种香豆素葡萄糖苷)。模式异形胞形成蓝细菌鱼腥藻PCC 7120对七叶苷的摄取受到α-葡萄糖苷蔗糖和麦芽糖的抑制。对突变体的分析鉴定出三种将七叶苷转运到细胞中的葡萄糖苷转运蛋白的组分:GlsC(Alr4781)和GlsP(All0261)分别是两种不同ABC转运蛋白的ATP结合亚基和通透酶亚基,而HepP(All1711)是一个主要协助转运蛋白超家族(MFS)蛋白,先前已证明其参与异形胞包膜的形成。葡萄糖苷转运蛋白基因的突变会损害鱼腥藻营养细胞之间荧光标记物(尤其是钙黄绿素)的转移。在细菌双杂交试验中,GlsP和HepP与隔膜连接相关蛋白SepJ相互作用,并且发现GlsC对于正常数量的隔膜肽聚糖纳米孔的形成以及SepJ的正常亚细胞定位是必需的。因此,除了它们在鱼腥藻营养摄取中可能发挥的作用外,葡萄糖苷转运蛋白还影响隔膜连接的结构和功能。形成异形胞的蓝细菌有能力进行有氧光合作用并同化大气中的二氧化碳和氮气。这些生物体以丝状体形式生长,分别在营养细胞和异形胞中特异性地固定这些气体。为了使丝状体生长,这些类型的细胞交换营养物质,包括蔗糖,蔗糖作为还原力和碳骨架的来源供异形胞使用。丝状体中细胞间蔗糖的移动通过隔膜连接进行,并且已用可被细胞摄取的荧光蔗糖类似物七叶苷进行了追踪。在这里,我们鉴定出鱼腥藻的α-葡萄糖苷转运蛋白,它们介导七叶苷的摄取,并且特别地,影响隔膜结构和隔膜连接的功能。

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