A study of the proteomic response of bat (<em>Pteropus alecto</em>) kidney cells to a viral mimic using iTRAQ and DIGE. — ASN Events
  1. Schedule
  2. Session 10: Poster Session C
  3. A study of the proteomic response of bat (Pteropus alecto) kidney cells to a viral mimic using iTRAQ and DIGE.

A study of the proteomic response of bat (Pteropus alecto) kidney cells to a viral mimic using iTRAQ and DIGE. (#390)

Lawrence Mok 1 2 , James Wynne 1 , Kris Ford 3 , Brian Shiell 1 , Tony Bacic 2 3 , Wojtek P Michalski 1
  1. CSIRO, Australian Animal Health Laboratory, East Geelong, Victoria, Australia
  2. Bio21 Institute for Molecular Science and Biotechnology, The University of Melbourne, Melbourne, Victoria, Australia
  3. ARC Centre of Excellence in Plant Cell Walls School of Botany , The University of Melbourne , Melbourne, Victoria, Australia

Bats have been identified as reservoirs for a number of zoonotic viruses including Ebola virus, Hendra virus, SARS coronavirus and Australian Bat Lyssavirus1.  Bats have the ability to transmit these viruses (a process known as spillover) to other susceptible species leading to severe illness and in some cases death. While these bat borne viruses may cause significant disease in their spillover host, bats remain asymptomatic and rarely shown any clinical signs of disease. The molecular mechanism by which bats control viral infection is therefore of significant value but is currently unknown. Here we utilise a model in vitro system and two proteomic techniques to characterise the response of bat cells to investigate the viral mimic, polyinosinic:polycytidylic acid (Poly I:C).

Poly I:C is an analog to dsRNA and is a mimic of virus infection, proven to induce an antiviral response in cells2. The cells used were immortalised bat kidney cells from Pteropus alecto (PaKiT03). P. alecto isa fruit bat native to Australia and also an asymptomatic carrier of the Hendra virus.

PaKiT03 cells were transfected with Poly I:C and sampled at 3 different time points post transfection. These samples were then analysed with 2D Differential In-Gel Electrophoresis (DIGE) and Isobaric Tag for Relative and Absolute Quantitation (iTRAQ) proteomics methods. Differentially expressed proteins in P. alecto cells following Poly I:C transfection can be used to compare with the type of known proteins that are expressed during live virus infection.

Each method identified a number of differentially expressed proteins, with some proteins being present in both datasets. Proteins were assessed as being differentially expressed in DIGE if they had a p-value of <.05. The cut off for differential expression in iTRAQ was a 1.3 fold change with less than 15% standard error.

The comparison of DIGE and iTRAQ techniques will help to assess which technique is more suitable for future proteomic analysis on P. alecto cells infected with live viruses.

  1. Calisher CH, Childs JE, Field HE, Holmes KV, Schountz T: Bats: important reservoir hosts of emerging viruses. Clin Microbiol Rev 2006, 19:531-545.
  2. Alexopoulou L, Holt AC, Medzhitov R, Flavell RA: Recognition of double-stranded RNA and activation of NF-kappaB by Toll-like receptor 3. Nature 2001, 413:732-738.