Mlouka Alyssa, Comte Katia, Castets Anne-Marie, Bouchier Christiane, Tandeau de Marsac Nicole
Unité des Cyanobactéries (URA-CNRS 2172), Département de Microbiologie fondamentale et médicale, Institut Pasteur, 75724 Paris Cedex 15, France.
J Bacteriol. 2004 Apr;186(8):2355-65. doi: 10.1128/JB.186.8.2355-2365.2004.
Microcystis aeruginosa is a planktonic unicellular cyanobacterium often responsible for seasonal mass occurrences at the surface of freshwater environments. An abundant production of intracellular structures, the gas vesicles, provides cells with buoyancy. A 8.7-kb gene cluster that comprises twelve genes involved in gas vesicle synthesis was identified. Ten of these are organized in two operons, gvpA(I)A(II)A(III)CNJX and gvpKFG, and two, gvpV and gvpW, are individually expressed. In an attempt to elucidate the basis for the frequent occurrence of nonbuoyant mutants in laboratory cultures, four gas vesicle-deficient mutants from two strains of M. aeruginosa, PCC 7806 and PCC 9354, were isolated and characterized. Their molecular analysis unveiled DNA rearrangements due to four different insertion elements that interrupted gvpN, gvpV, or gvpW or led to the deletion of the gvpA(I)-A(III) region. While gvpA, encoding the major gas vesicle structural protein, was expressed in the gvpN, gvpV, and gvpW mutants, immunodetection revealed no corresponding GvpA protein. Moreover, the absence of a gas vesicle structure was confirmed by electron microscopy. This study brings out clues concerning the process driving loss of buoyancy in M. aeruginosa and reveals the requirement for gas vesicle synthesis of two newly described genes, gvpV and gvpW.
铜绿微囊藻是一种浮游单细胞蓝细菌,常导致淡水环境表面季节性大量出现。细胞内大量产生的气体囊泡为细胞提供浮力。鉴定出一个8.7 kb的基因簇,其中包含12个参与气体囊泡合成的基因。其中10个基因组成两个操纵子,即gvpA(I)A(II)A(III)CNJX和gvpKFG,另外两个基因gvpV和gvpW单独表达。为了阐明实验室培养物中无浮力突变体频繁出现的原因,从铜绿微囊藻的两个菌株PCC 7806和PCC 9354中分离并鉴定了四个气体囊泡缺陷突变体。它们的分子分析揭示了由于四种不同的插入元件导致的DNA重排,这些元件中断了gvpN、gvpV或gvpW,或导致gvpA(I)-A(III)区域的缺失。虽然编码主要气体囊泡结构蛋白的gvpA在gvpN、gvpV和gvpW突变体中表达,但免疫检测未发现相应的GvpA蛋白。此外,电子显微镜证实没有气体囊泡结构。这项研究揭示了有关铜绿微囊藻浮力丧失过程的线索,并揭示了两个新描述的基因gvpV和gvpW对气体囊泡合成的需求。