PLoS One. 2012; 7(3): e33408
Holmes A, Lindestam Arlehamn CS, Wang D, Mitchell TJ, Evans TJ, Roe AJ
E. coli O157 carries two genes encoding a effector proteins NleH1 as well as NleH2 which are 87% identical. Despite a similarity between a proteins, a promoter regions upstream of a genes encoding a effectors are more anomalous suggesting that a actual countenance of a genes might be differentially regulated. This was tested by formulating reporter fusions as well as examining their countenance in different genetic backgrounds, media as well as on hit with horde cells. The function of a proteins was also tested following transfection into horde cells.Expression of both NleH1 as well as NleH2 was enhanced when cultured underneath conditions that stimulated countenance of a Type Three Secretion System (T3SS) as well as was influenced by a regulators Ler as well as GrlA. Maximal countenance of NleH1 required 531 bp of a upstream untranslated region though NleH2 required only 113 bp. Interestingly, hit with horde cells strongly repressed countenance of both NleH1 as well as NleH2. Following transfection, both proteins constructed only teenager effects on NF-B activation when assessed using a NF-B luciferase reporter assay, a result that is consistent with a recent report demonstrating a dependence on RPS3 for NleH1 modulation of NF-B.This study demonstrates a importance of considering gene regulation when studying bacterial effector proteins. Despite their sequence similarity, NleH1 as well as NleH2 are expressed differentially as well as may, therefore, be translocated at distinct times during an infection.
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