Proteomics

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Combining transcriptome and proteome analysis to identify and characterize the RpoS regulon in Pseudomonas fluorescens


ABSTRACT: Background: Microorganisms are the major cause of food spoilage during storage, processing and distribution. Pseudomonas fluorescens is a typical spoilage bacterium that contributes to a large extent to the spoilage process of proteinaceous food. RpoS is considered an important global regulator involved in stress survival and virulence in many pathogens. Our previous work revealed that RpoS contributed to the spoilage activities of P. fluorescens by regulating resistance to different stress conditions, extracellular acylated homoserine lactone (AHL) levels, extracellular protease and total volatile basic nitrogen (TVB-N) production. However, RpoS-dependent genes in P. fluorescens remained undefined. Results: RNA-seq transcriptomics analysis combined with quantitative proteomics analysis basing on multiplexed isobaric tandem mass tag (TMT) labeling was performed for the P. fluorescens wild-type strain UK4 and its derivative carrying a rpoS mutation. A total of 375 differentially expressed genes (DEGs) and 212 differentially expressed proteins (DEPs) were identified in these two backgrounds. The DGEs were further verified by qRT-PCR tests, and the genes directly regulated by RpoS were confirmed by 5’-RACE-PCR sequencing. The combining transcriptome and proteome analysis revealed a role of this regulator in several cellular processes, including polysaccharide metabolism, intracellular secretion and extracellular structures, cell well biogenesis, stress responses, ammonia and biogenic amine production, which may contribute to biofilm formation, stress resistance and spoilage activities of P. fluorescens. Moreover, in this work we indeed observed that RpoS contributed to the production of the macrocolony biofilm’s matrix.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Pseudomonas Fluorescens

TISSUE(S): Whole Body

SUBMITTER: xiaonan fu  

LAB HEAD: Xiaoxiang Liu

PROVIDER: PXD010845 | Pride | 2019-11-12

REPOSITORIES: Pride

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Publications

Combined Transcriptome and Proteome Analysis of RpoS Regulon Reveals Its Role in Spoilage Potential of <i>Pseudomonas fluorescens</i>.

Liu Xiaoxiang X   Xu Jun J   Zhu Junli J   Du Peng P   Sun Aihua A  

Frontiers in microbiology 20190206


Microbial contamination is considered the main cause of food spoilage. <i>Pseudomonas fluorescens</i> is a typical spoilage bacterium contributing to a large extent to the spoilage process of proteinaceous foods. RpoS is known as an alternative sigma factor controlling stress resistance and virulence in many pathogens. Our previous work revealed that RpoS contributes to the spoilage activities of <i>P</i>. <i>fluorescens</i> by regulating resistance to different stress conditions, extracellular  ...[more]

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