Identification and initial characterisation of a protein involved in Campylobacter jejuni cell shape.
Identification and initial characterisation of a protein involved in Campylobacter jejuni cell shape.
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DOI：
10.1016/j.micpath.2017.01.042
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发表时间：
2017-03
影响因子：
3.8
通讯作者：
Grant AJ
中科院分区：
文献类型：
--
作者：
Esson D;Gupta S;Bailey D;Wigley P;Wedley A;Mather AE;Méric G;Mastroeni P;Sheppard SK;Thomson NR;Parkhill J;Maskell DJ;Christie G;Grant AJ
空肠弯曲杆菌是细菌性食源性疾病的主要原因。虽然螺旋细胞形状被认为对空肠梭菌的发病机制很重要，但这种细菌能够采用其他形态。为了更好地了解螺旋形空肠梭菌是如何保持其形状的，从而了解任何相关的定植、致病性或其他优势，首先重要的是确定相关的基因和蛋白质。到目前为止，两种肽聚糖修饰酶ppgp1和ppgp2已被证明是空肠梭菌螺旋细胞形状所必需的。我们对空肠梭菌的约2000个转座子突变体进行了细胞形状突变的视觉筛选。细胞形状改变突变体、定向突变体、野生型种群和分离的螺旋形和杆状“野生型”空肠C.的全基因组序列数据发现，pgp1和pgp2发生了许多不同的突变，导致螺旋形到杆状细菌细胞形状的变化。我们还确定了一个曲率损失的孤立体。在本研究中，我们确定了该分离物的基因组变化，并发现有针对性地删除该变化的基因导致细菌失去曲率。通过提供反式基因，使细胞恢复螺旋形。我们研究了基因缺失对细菌运动、组织培养细胞的粘附和入侵以及鸡定植的影响，以及对肽聚糖小囊多肽谱的影响。我们的工作确定了另一个与螺旋细胞形状有关的因素。鉴定出一株具有曲率损失的分离空肠梭菌。建立了81-176中CJJ81176_1105和M1中CJM1_1064的靶向缺失基因。确定的CJJ81176_1105和CJM1_1064基因缺失突变改变了空肠梭菌的运动和与Caco-2细胞的相互作用。确定的CJM1_1064基因缺失突变体不改变鸡空肠梭菌的定殖。
Campylobacter jejuni is the leading cause of bacterial food borne illness. While helical cell shape is considered important for C. jejuni pathogenesis, this bacterium is capable of adopting other morphologies. To better understand how helical-shaped C. jejuni maintain their shape and thus any associated colonisation, pathogenicity or other advantage, it is first important to identify the genes and proteins involved. So far, two peptidoglycan modifying enzymes Pgp1 and Pgp2 have been shown to be required for C. jejuni helical cell shape. We performed a visual screen of ∼2000 transposon mutants of C. jejuni for cell shape mutants. Whole genome sequence data of the mutants with altered cell shape, directed mutants, wild type stocks and isolated helical and rod-shaped ‘wild type’ C. jejuni, identified a number of different mutations in pgp1 and pgp2, which result in a change in helical to rod bacterial cell shape. We also identified an isolate with a loss of curvature. In this study, we have identified the genomic change in this isolate, and found that targeted deletion of the gene with the change resulted in bacteria with loss of curvature. Helical cell shape was restored by supplying the gene in trans. We examined the effect of loss of the gene on bacterial motility, adhesion and invasion of tissue culture cells and chicken colonisation, as well as the effect on the muropeptide profile of the peptidoglycan sacculus. Our work identifies another factor involved in helical cell shape. A C. jejuni isolate with a loss of curvature was identified. A targeted gene deletion of CJJ81176_1105 in 81–176 and CJM1_1064 in M1 were created. Defined gene deletion mutants of CJJ81176_1105 and CJM1_1064 alter C. jejuni motility and interaction with Caco-2 cells. Defined gene deletion mutant of CJM1_1064 does not alter C. jejuni colonisation of chickens.