Identification of AbrB-regulated genes involved in biofilm formation by Bacillus subtilis
Identification of AbrB-regulated genes involved in biofilm formation by Bacillus subtilis
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DOI：
10.1111/j.1365-2958.2004.04023.x
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发表时间：
2004-05-01
影响因子：
3.6
通讯作者：
Lazazzera, BA
中科院分区：
文献类型：
--
作者：
Hamon, MA;Stanley, NR;Lazazzera, BA
枯草芽孢杆菌是一种普遍存在的土壤细菌，其在由转录因子AbrB负控制的过程中形成生物膜。鉴定B生物膜形成所需的AbrB调控基因。枯草芽孢杆菌中，进行spo 0A abrB和sigH abrB突变菌株形成的生物膜的全基因组表达谱研究。这些数据，与先前发表的spo 0A和sigH突变株的DNA微阵列分析相一致，导致鉴定出39个似乎被AbrB抑制的操纵子。这些操纵子中的8个先前已被证明被AbrB抑制，我们通过逆转录-PCR证实了AbrB对另外6个操纵子的抑制。在这项研究中确定的AbrB抑制基因参与已知由AbrB调节的过程，如细胞外降解酶的产生和氨基酸代谢，以及以前不知道由AbrB调节的过程，如膜生物能学和细胞壁功能。为了确定这些AbrB调节基因中的任何一个是否在生物膜形成中起作用，我们测试了23种突变体，每种突变体在不同的AbrB调节操纵子中具有破坏，以形成生物膜的能力。两个突变体在生物膜形成中具有大于两倍的缺陷。yoaW突变体表现出深度减少的生物膜结构，而sipW突变体仅表现出表面附着的细胞，不形成成熟的生物膜。YoaW是一种推定的分泌蛋白，SipW是一种信号肽酶。这是分泌蛋白在枯草芽孢杆菌生物膜形成中起作用的第一个证据。
Bacillus subtilis is a ubiquitous soil bacterium that forms biofilms in a process that is negatively controlled by the transcription factor AbrB. To identify the AbrB-regulated genes required for biofilm formation by B. subtilis, genome-wide expression profiling studies of biofilms formed by spo0A abrB and sigH abrB mutant strains were performed. These data, in concert with previously published DNA microarray analysis of spo0A and sigH mutant strains, led to the identification of 39 operons that appear to be repressed by AbrB. Eight of these operons had previously been shown to be repressed by AbrB, and we confirmed AbrB repression for a further six operons by reverse transcription-PCR. The AbrB-repressed genes identified in this study are involved in processes known to be regulated by AbrB, such as extracellular degradative enzyme production and amino acid metabolism, and processes not previously known to be regulated by AbrB, such as membrane bioenergetics and cell wall functions. To determine whether any of these AbrB-regulated genes had a role in biofilm formation, we tested 23 mutants, each with a disruption in a different AbrB-regulated operon, for the ability to form biofilms. Two mutants had a greater than twofold defect in biofilm formation. A yoaW mutant exhibited a biofilm structure with reduced depth, and a sipW mutant exhibited only surface-attached cells and did not form a mature biofilm. YoaW is a putative secreted protein, and SipW is a signal peptidase. This is the first evidence that secreted proteins have a role in biofilm formation by Bacillus subtilis.