肺炎链球菌中输出蛋白的基因鉴定

Genetic identification of exported proteins in Streptococcus pneumoniae.

作者信息

Pearce B J, Yin Y B, Masure H R

机构信息

Laboratory of Molecular Infectious Diseases, Rockefeller University, New York, New York 10021.

出版信息

Mol Microbiol. 1993 Sep;9(5):1037-50. doi: 10.1111/j.1365-2958.1993.tb01233.x.

DOI:10.1111/j.1365-2958.1993.tb01233.x

PMID:7934910

Abstract

A strategy was developed to mutate and genetically identify exported proteins in Streptococcus pneumoniae. Vectors were created and used to screen pneumococcal DNA in Escherichia coli and S. pneumoniae for translational gene fusions to alkaline phosphatase (PhoA). Twenty five PhoA+ pneumococcal mutants were isolated and the loci from eight of these mutants showed similarity to known exported or membrane-associated proteins. Homologues were found to: (i) protein-dependent peptide permeases, (ii) penicillin-binding proteins, (iii) Clp proteases, (iv) two-component sensor regulators, (v) the phosphoenolpyruvate: carbohydrate phosphotransferases permeases, (vi) membrane-associated dehydrogenases, (vii) P-type (E1E2-type) cation transport ATPases, (viii) ABC transporters responsible for the translocation of the RTX class of bacterial toxins. Unexpectedly one PhoA+ mutant contained a fusion to a member of the DEAD protein family of ATP-dependent RNA helicases suggesting export of these proteins.

摘要

已制定出一种策略，用于对肺炎链球菌中的输出蛋白进行突变和基因鉴定。构建了载体，并用于在大肠杆菌和肺炎链球菌中筛选肺炎球菌DNA，以寻找与碱性磷酸酶（PhoA）的翻译基因融合。分离出了25个PhoA +肺炎球菌突变体，其中8个突变体的基因座与已知的输出或膜相关蛋白具有相似性。发现的同源物有：（i）蛋白质依赖性肽通透酶，（ii）青霉素结合蛋白，（iii）Clp蛋白酶，（iv）双组分传感器调节因子，（v）磷酸烯醇丙酮酸：碳水化合物磷酸转移酶通透酶，（vi）膜相关脱氢酶，（vii）P型（E1E2型）阳离子转运ATP酶，（viii）负责RTX类细菌毒素转运的ABC转运蛋白。出乎意料的是，一个PhoA +突变体包含与ATP依赖性RNA解旋酶的DEAD蛋白家族成员的融合，这表明这些蛋白会输出。