title: A novel member of the let-7 microRNA family is associated with developmental transitions in filarial nematode parasites.
abstract: BACKGROUND
  Filarial nematodes are important pathogens in the tropics transmitted to humans via the bite of blood sucking arthropod vectors. The molecular mechanisms underpinning survival and differentiation of these parasites following transmission are poorly understood. microRNAs are small non-coding RNA molecules that regulate target mRNAs and we set out to investigate whether they play a role in the infection event.
  
  
  RESULTS
  microRNAs differentially expressed during the early post-infective stages of Brugia pahangi L3 were identified by microarray analysis. One of these, bpa-miR-5364, was selected for further study as it is upregulated ~12-fold at 24 hours post-infection, is specific to clade III nematodes, and is a novel member of the let-7 family, which are known to have key developmental functions in the free-living nematode Caenorhabditis elegans. Predicted mRNA targets of bpa-miR-5364 were identified using bioinformatics and comparative genomics approaches that relied on the conservation of miR-5364 binding sites in the orthologous mRNAs of other filarial nematodes. Finally, we confirmed the interaction between bpa-miR-5364 and three of its predicted targets using a dual luciferase assay.
  
  
  CONCLUSIONS
  These data provide new insight into the molecular mechanisms underpinning the transmission of third stage larvae of filarial nematodes from vector to mammal. This study is the first to identify parasitic nematode mRNAs that are verified targets of specific microRNAs and demonstrates that post-transcriptional control of gene expression via stage-specific expression of microRNAs may be important in the success of filarial infection.
authors.affiliation: Institute of Biodiversity, Animal Health and Comparative Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, Garscube Estate, Bearsden Road, Glasgow, G61 1QH, UK. alan.winter@glasgow.ac.uk.
authors.forename: Alan D
authors.initials: AD
authors.lastname: Winter
authors.affiliation: Institute of Biodiversity, Animal Health and Comparative Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, Garscube Estate, Bearsden Road, Glasgow, G61 1QH, UK. victoria.gillan@glasgow.ac.uk.
authors.forename: Victoria
authors.initials: V
authors.lastname: Gillan
authors.affiliation: Institute of Biodiversity, Animal Health and Comparative Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, Garscube Estate, Bearsden Road, Glasgow, G61 1QH, UK. kirsty.maitland@glasgow.ac.uk.
authors.forename: Kirsty
authors.initials: K
authors.lastname: Maitland
authors.affiliation: School of Veterinary Medicine and Science, University of Nottingham, Sutton Bonington Campus, Leicestershire, LE12 5RD, UK. richard.emes@nottingham.ac.uk.|Advanced Data Analysis Centre, University of Nottingham, Nottingham, UK. richard.emes@nottingham.ac.uk.
authors.forename: Richard D
authors.initials: RD
authors.lastname: Emes
authors.affiliation: Institute of Biodiversity, Animal Health and Comparative Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, Garscube Estate, Bearsden Road, Glasgow, G61 1QH, UK. brett.roberts@glasgow.ac.uk.
authors.forename: Brett
authors.initials: B
authors.lastname: Roberts
authors.affiliation: Institute of Biodiversity, Animal Health and Comparative Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, Garscube Estate, Bearsden Road, Glasgow, G61 1QH, UK. gillian.mccormack@glasgow.ac.uk.
authors.forename: Gillian
authors.initials: G
authors.lastname: McCormack
authors.affiliation: Institute of Biodiversity, Animal Health and Comparative Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, Garscube Estate, Bearsden Road, Glasgow, G61 1QH, UK. willie.weir@glasgow.ac.uk.
authors.forename: William
authors.initials: W
authors.lastname: Weir
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridgeshire, CB10 1SA, UK. ap6@sanger.ac.uk.
authors.forename: Anna V
authors.initials: AV
authors.lastname: Protasio
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridgeshire, CB10 1SA, UK. neh@sanger.ac.uk.
authors.forename: Nancy
authors.initials: N
authors.lastname: Holroyd
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridgeshire, CB10 1SA, UK. mb4@sanger.ac.uk.
authors.forename: Matthew
authors.initials: M
authors.lastname: Berriman
authors.affiliation: Institute of Biodiversity, Animal Health and Comparative Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, Garscube Estate, Bearsden Road, Glasgow, G61 1QH, UK. collette.britton@glasgow.ac.uk.
authors.forename: Collette
authors.initials: C
authors.lastname: Britton
authors.affiliation: Institute of Biodiversity, Animal Health and Comparative Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, Garscube Estate, Bearsden Road, Glasgow, G61 1QH, UK. eileen.devaney@glasgow.ac.uk.
authors.forename: Eileen
authors.initials: E
authors.lastname: Devaney
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D001483:Base Sequence; D017179:Brugia pahangi / Q000145:classification / Q000235:genetics* / Q000254:growth & development; D019295:Computational Biology; D005260:Female; D008018:Life Cycle Stages / Q000235:genetics; D008297:Male; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D016376:Oligonucleotides, Antisense / Q000378:metabolism; D010802:Phylogeny; D012333:RNA, Messenger / Q000037:antagonists & inhibitors / Q000378:metabolism; D016415:Sequence Alignment; D017423:Sequence Analysis, RNA; D059467:Transcriptome
chemical_list: D020413:3' Untranslated Regions; D035683:MicroRNAs; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger