title: Quantitative Analysis of MicroRNAs in Vaccinia virus Infection Reveals Diversity in Their Susceptibility to Modification and Suppression.
abstract: Vaccinia virus (VACV) is a large cytoplasmic DNA virus that causes dramatic alterations to many cellular pathways including microRNA biogenesis. The virus encodes a poly(A) polymerase which was previously shown to add poly(A) tails to the 3' end of cellular miRNAs, resulting in their degradation by 24 hours post infection (hpi). Here we used small RNA sequencing to quantify the impact of VACV infection on cellular miRNAs in human cells at both early (6 h) and late (24 h) times post infection. A detailed quantitative analysis of individual miRNAs revealed marked diversity in the extent of their modification and relative change in abundance during infection. Some miRNAs became highly modified (e.g. miR-29a-3p, miR-27b-3p) whereas others appeared resistant (e.g. miR-16-5p). Furthermore, miRNAs that were highly tailed at 6 hpi were not necessarily among the most reduced at 24 hpi. These results suggest that intrinsic features of human cellular miRNAs cause them to be differentially polyadenylated and altered in abundance during VACV infection. We also demonstrate that intermediate and late VACV gene expression are required for optimal repression of some miRNAs including miR-27-3p. Overall this work reveals complex and varied consequences of VACV infection on host miRNAs and identifies miRNAs which are largely resistant to VACV-induced polyadenylation and are therefore present at functional levels during the initial stages of infection and replication.
authors.affiliation: Centre for Immunity, Infection and Evolution, University of Edinburgh, King's Buildings, Edinburgh, United Kingdom.
authors.forename: Amy H
authors.initials: AH
authors.lastname: Buck
authors.affiliation: Centre for Immunity, Infection and Evolution, University of Edinburgh, King's Buildings, Edinburgh, United Kingdom.
authors.forename: Alasdair
authors.initials: A
authors.lastname: Ivens
authors.affiliation: Centre for Immunity, Infection and Evolution, University of Edinburgh, King's Buildings, Edinburgh, United Kingdom.
authors.forename: Katrina
authors.initials: K
authors.lastname: Gordon
authors.affiliation: Infection and Immunity, The Roslin Institute / Royal (Dick) School of Veterinary Studies, Easter Bush, Midlothian, United Kingdom.
authors.forename: Nicola
authors.initials: N
authors.lastname: Craig
authors.affiliation: Infection and Immunity, The Roslin Institute / Royal (Dick) School of Veterinary Studies, Easter Bush, Midlothian, United Kingdom.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Houzelle
authors.affiliation: Infection and Immunity, The Roslin Institute / Royal (Dick) School of Veterinary Studies, Easter Bush, Midlothian, United Kingdom.
authors.forename: Alice
authors.initials: A
authors.lastname: Roche
authors.affiliation: Infection and Immunity, The Roslin Institute / Royal (Dick) School of Veterinary Studies, Easter Bush, Midlothian, United Kingdom.
authors.forename: Neil
authors.initials: N
authors.lastname: Turnbull
authors.affiliation: Infection and Immunity, The Roslin Institute / Royal (Dick) School of Veterinary Studies, Easter Bush, Midlothian, United Kingdom.
authors.forename: Philippa M
authors.initials: PM
authors.lastname: Beard
mesh_terms: D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D020869:Gene Expression Profiling / Q000379:methods*; D020022:Genetic Predisposition to Disease / Q000235:genetics; D014644:Genetic Variation; D006367:HeLa Cells; D006801:Humans; D035683:MicroRNAs / Q000737:chemistry / Q000235:genetics*; D017423:Sequence Analysis, RNA / Q000379:methods*; D014615:Vaccinia / Q000235:genetics / Q000821:virology; D014616:Vaccinia virus / Q000502:physiology*
chemical_list: D035683:MicroRNAs