title: Twin-arginine signal peptide of Bacillus subtilis YwbN can direct Tat-dependent secretion of methyl parathion hydrolase.
abstract: The twin-arginine translocation (Tat) pathway exports folded proteins across the cytoplasmic membranes of bacteria and archaea. Two parallel Tat pathways (TatAdCd and TatAyCy systems) with distinct substrate specificities have previously been discovered in Bacillus subtilis. In this study, to secrete methyl parathion hydrolase (MPH) into the growth medium, the twin-arginine signal peptide of B. subtilis YwbN was used to target MPH to the Tat pathway of B. subtilis. Western blot analysis and MPH assays demonstrated that active MPH was secreted into the culture supernatant of wild-type cells. No MPH secretion occurred in a total-tat2 mutant, indicating that the observed export in wild-type cells was mediated exclusively by the Tat pathway. Export was fully blocked in a tatAyCy mutant. In contrast, the tatAdCd mutant was still capable of secreting MPH. These results indicated that the MPH secretion directed by the YwbN signal peptide was specifically mediated by the TatAyCy system. The N-terminal sequence of secreted MPH was determined as AAPQVR, demonstrating that the YwbN signal peptide had been processed correctly. This is the first report of functional secretion of a heterologous protein via the B. subtilis TatAyCy system. This study highlights the potential of the TatAyCy system to be used for secretion of other heterologous proteins in B. subtilis.
authors.affiliation: College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, §Key Laboratory of Molecular Microbiology and Technology for Ministry of Education, Department of Microbiology, College of Life Sciences, and ¶Key Laboratory of Pollution Processes and Environmental Criteria for Ministry of Education, College of Environmental Science and Engineering, Nankai University , Tianjin 300071, People's Republic of China.
authors.forename: Ruihua
authors.initials: R
authors.lastname: Liu
authors.forename: Zhenqiang
authors.initials: Z
authors.lastname: Zuo
authors.forename: Yingming
authors.initials: Y
authors.lastname: Xu
authors.forename: Cunjiang
authors.initials: C
authors.lastname: Song
authors.forename: Hong
authors.initials: H
authors.lastname: Jiang
authors.forename: Chuanling
authors.initials: C
authors.lastname: Qiao
authors.forename: Ping
authors.initials: P
authors.lastname: Xu
authors.forename: Qixing
authors.initials: Q
authors.lastname: Zhou
authors.forename: Chao
authors.initials: C
authors.lastname: Yang
mesh_terms: D001412:Bacillus subtilis / Q000737:chemistry / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D058947:Bacterial Secretion Systems; D001483:Base Sequence; D008969:Molecular Sequence Data; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism*; D021382:Protein Sorting Signals; D021381:Protein Transport
chemical_list: D001426:Bacterial Proteins; D058947:Bacterial Secretion Systems; D021382:Protein Sorting Signals; D010744:Phosphoric Monoester Hydrolases