Bucarey Sergio A, Villagra Nicolas A, Fuentes Juan A, Mora Guido C
Programa Doctorado de Genética Molecular y Microbiología, Pontificia Universidad Católica de Chile, Santiago, Chile.
Genetics. 2006 May;173(1):25-34. doi: 10.1534/genetics.105.054700. Epub 2006 Feb 19.
The Salmonella enterica tsx gene encodes a nucleoside-specific outer membrane channel. The Tsx porin is essential for the prototrophic growth of S. enterica sv. Typhi in the absence of nucleosides. RT-PCR analysis shows that the tsx gene is cotranscribed with an open reading frame unique to S. enterica, impX (STY0450), which encodes an inner membrane protein 108 amino acids in length, which is predicted to have only two transmembrane alpha-helices. Fusions of the lacZ gene to both tsx and impX reveal that the transcription of both genes is induced in the presence of adenosine. A null mutation in the S. Typhi impX gene suppresses the induced auxotrophy for adenosine or thymidine resulting from a tsx mutation and confers sensitivity to high concentrations of adenosine or thymidine. The ImpX protein, when tagged with a 3xFLAG epitope, is functional and associates with the inner membrane; impX mutants are defective in the export of 3H-radiolabeled thymidine. Taken together, these and other results suggest that the S. Typhi Tsx porin and ImpX inner membrane protein facilitate competing mechanisms of thymidine influx and efflux, respectively, to maintain the steady-state levels of internal nucleoside pools.
肠炎沙门氏菌tsx基因编码一种核苷特异性外膜通道。在缺乏核苷的情况下,Tsx孔蛋白对于肠炎沙门氏菌伤寒亚种的原养型生长至关重要。逆转录聚合酶链反应(RT-PCR)分析表明,tsx基因与肠炎沙门氏菌特有的一个开放阅读框impX(STY0450)共转录,impX编码一种长度为108个氨基酸的内膜蛋白,预计该蛋白仅具有两个跨膜α螺旋。将lacZ基因与tsx和impX融合表明,这两个基因的转录在腺苷存在的情况下被诱导。伤寒沙门氏菌impX基因的无效突变抑制了由tsx突变导致的对腺苷或胸苷的诱导性营养缺陷,并赋予对高浓度腺苷或胸苷的敏感性。当用3xFLAG表位标记时,ImpX蛋白具有功能并与内膜相关联;impX突变体在3H放射性标记胸苷的输出方面存在缺陷。综上所述,这些以及其他结果表明,伤寒沙门氏菌Tsx孔蛋白和ImpX内膜蛋白分别促进了胸苷流入和流出的竞争机制,以维持内部核苷池的稳态水平。