title: Suppression of fabB Mutation by fabF1 Is Mediated by Transcription Read-through in Shewanella oneidensis.
abstract: As type II fatty acid synthesis is essential for the growth of Escherichia coli, its many components are regarded as potential targets for novel antibacterial drugs. Among them, β-ketoacyl-acyl carrier protein (ACP) synthase (KAS) FabB is the exclusive factor for elongation of the cis-3-decenoyl-ACP (cis-3-C10-ACP). In our previous study, we presented evidence to suggest that this may not be the case in Shewanella oneidensis, an emerging model gammaproteobacterium renowned for its respiratory versatility. Here, we identified FabF1, another KAS, as a functional replacement for FabB in S. oneidensis In fabB+or desA+(encoding a desaturase) cells, which are capable of making unsaturated fatty acids (UFA), FabF1 is barely produced. However, UFA auxotroph mutants devoid of both fabB and desA genes can be spontaneously converted to suppressor strains, which no longer require exogenous UFAs for growth. Suppression is caused by a TGTTTT deletion in the region upstream of the fabF1 gene, resulting in enhanced FabF1 production. We further demonstrated that the deletion leads to transcription read-through of the terminator for acpP, an acyl carrier protein gene immediately upstream of fabF1 There are multiple tandem repeats in the region covering the terminator, and the TGTTTT deletion, as well as others, compromises the terminator efficacy. In addition, FabF2 also shows an ability to complement the FabB loss, albeit substantially less effectively than FabF1.
  
  
  
  It has been firmly established that FabB for UFA synthesis via type II fatty acid synthesis in FabA-containing bacteria such as E. coli is essential. However, S. oneidensis appears to be an exception. In this bacterium, FabF1, when sufficiently expressed, is able to fully complement the FabB loss. Importantly, such a capability can be obtained by spontaneous mutations, which lead to transcription read-through. Therefore, our data, by identifying the functional overlap between FabB and FabFs, provide new insights into the current understanding of KAS and help reveal novel ways to block UFA synthesis for therapeutic purposes.
authors: Institute of Microbiology and College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, China.
authors: Meng
authors: M
authors: Li
authors: Institute of Microbiology and College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, China.
authors: Qiu
authors: Q
authors: Meng
authors: Institute of Microbiology and College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, China.
authors: Huihui
authors: H
authors: Fu
authors: Institute of Microbiology and College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, China.
authors: Qixia
authors: Q
authors: Luo
authors: Institute of Microbiology and College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, China haichung@zju.edu.cn.
authors: Haichun
authors: H
authors: Gao
mesh_terms: D015099:3-Oxoacyl-(Acyl-Carrier-Protein) Synthase / Q000235:genetics* / Q000378:metabolism; D000123:Acetyltransferases / Q000235:genetics / Q000378:metabolism; D000213:Acyl Carrier Protein / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism; D054889:Fatty Acid Synthase, Type II / Q000235:genetics* / Q000378:metabolism; D005231:Fatty Acids, Unsaturated / Q000096:biosynthesis*; D015964:Gene Expression Regulation, Bacterial; D055879:Gene Knock-In Techniques; D017384:Sequence Deletion; D020592:Shewanella / Q000235:genetics* / Q000378:metabolism; D024363:Transcription Initiation Site
chemical_list: D000213:Acyl Carrier Protein; D029968:Escherichia coli Proteins; D005231:Fatty Acids, Unsaturated; D000123:Acetyltransferases; C557137:fabF protein, E coli; D015099:3-Oxoacyl-(Acyl-Carrier-Protein) Synthase; C490369:FabB protein, E coli; D054889:Fatty Acid Synthase, Type II