title: Uropathogenic Escherichia coli Releases Extracellular Vesicles That Are Associated with RNA.
abstract: BACKGROUND
  Bacterium-to-host signalling during infection is a complex process involving proteins, lipids and other diffusible signals that manipulate host cell biology for pathogen survival. Bacteria also release membrane vesicles (MV) that can carry a cargo of effector molecules directly into host cells. Supported by recent publications, we hypothesised that these MVs also associate with RNA, which may be directly involved in the modulation of the host response to infection.
  
  
  RESULTS
  Using the uropathogenic Escherichia coli (UPEC) strain 536, we have isolated MVs and found they carry a range of RNA species. Density gradient centrifugation further fractionated and characterised the MV preparation and confirmed that the isolated RNA was associated with the highest particle and protein containing fractions. Using a new approach, RNA-sequencing of libraries derived from three different 'size' RNA populations (<50nt, 50-200nt and 200nt+) isolated from MVs has enabled us to now report the first example of a complete bacterial MV-RNA profile. These data show that MVs carry rRNA, tRNAs, other small RNAs as well as full-length protein coding mRNAs. Confocal microscopy visualised the delivery of lipid labelled MVs into cultured bladder epithelial cells and showed their RNA cargo labelled with 5-EU (5-ethynyl uridine), was transported into the host cell cytoplasm and nucleus. MV RNA uptake by the cells was confirmed by droplet digital RT-PCR of csrC. It was estimated that 1% of MV RNA cargo is delivered into cultured cells.
  
  
  CONCLUSIONS
  These data add to the growing evidence of pathogenic bacterial MV being associated a wide range of RNAs. It further raises the plausibility for MV-RNA-mediated cross-kingdom communication whereby they influence host cell function during the infection process.
authors: Department of Molecular Medicine and Pathology, University of Auckland, Auckland, New Zealand.|Department of Surgery, University of Auckland, Auckland, New Zealand.
authors: Cherie
authors: C
authors: Blenkiron
authors: Department of Molecular Medicine and Pathology, University of Auckland, Auckland, New Zealand.|Department of Surgery, University of Auckland, Auckland, New Zealand.
authors: Denis
authors: D
authors: Simonov
authors: Department of Obstetrics and Gynaecology, University of Auckland, Auckland, New Zealand.
authors: Anita
authors: A
authors: Muthukaruppan
authors: Bioinformatics Institute, University of Auckland, Auckland, New Zealand.
authors: Peter
authors: P
authors: Tsai
authors: Department of Molecular Medicine and Pathology, University of Auckland, Auckland, New Zealand.
authors: Priscila
authors: P
authors: Dauros
authors: Department of Molecular Medicine and Pathology, University of Auckland, Auckland, New Zealand.
authors: Sasha
authors: S
authors: Green
authors: Department of Surgery, University of Auckland, Auckland, New Zealand.|School of Biological Sciences, University of Auckland, Auckland, New Zealand.
authors: Jiwon
authors: J
authors: Hong
authors: Department of Molecular Medicine and Pathology, University of Auckland, Auckland, New Zealand.|Bioinformatics Institute, University of Auckland, Auckland, New Zealand.|Maurice Wilkins Centre, University of Auckland, Auckland, New Zealand.
authors: Cristin G
authors: CG
authors: Print
authors: Department of Molecular Medicine and Pathology, University of Auckland, Auckland, New Zealand.
authors: Simon
authors: S
authors: Swift
authors: Department of Surgery, University of Auckland, Auckland, New Zealand.|School of Biological Sciences, University of Auckland, Auckland, New Zealand.|Maurice Wilkins Centre, University of Auckland, Auckland, New Zealand.
authors: Anthony R
authors: AR
authors: Phillips
mesh_terms: D045744:Cell Line, Tumor; D002499:Centrifugation, Density Gradient; D004847:Epithelial Cells / Q000382:microbiology / Q000473:pathology; D004927:Escherichia coli Infections / Q000382:microbiology; D000067128:Extracellular Vesicles / Q000235:genetics*; D006801:Humans; D012329:RNA, Bacterial / Q000235:genetics / Q000378:metabolism; D012335:RNA, Ribosomal / Q000235:genetics / Q000378:metabolism; D012343:RNA, Transfer / Q000235:genetics / Q000378:metabolism; D001749:Urinary Bladder Neoplasms / Q000382:microbiology; D056689:Uropathogenic Escherichia coli / Q000166:cytology / Q000235:genetics* / Q000472:pathogenicity
chemical_list: D012329:RNA, Bacterial; D012335:RNA, Ribosomal; D012343:RNA, Transfer