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Characterization of the Interferon Alpha inducible protein LY6E

Friday, November 08, 2013 — Poster Session IV

2:00 p.m. – 4:00 p.m.

FAES Academic Center (Upper-Level Terrace)

NIAID

IMMUNO-7

Authors

  • DS Green
  • J Bekiscz
  • M Dolan
  • KC Zoon

Abstract

Ly6E is a member of the Ly family of proteins whose genes are clustered on Chromosome 8. In micro array experiments performed by both our lab and others has shown the Ly6E is highly up regulated in response to Interferon Alpha (IFNa). Despite the identification of Ly6E as an IFNa inducible gene, the protein has not been characterized. Using computer modeling we propose that Ly6E is a spherical protein that is connected to the cell membrane through a GPI anchor. Through the modeling and sequence alignment we created a rabbit monoclonal antibody to a unique region of Ly6E. We validated the antibody both through Western Blot (WB) and Flow Cytometric (FC) analysis. Using FC and WB we found that the cell line A549 expressed Ly6E constitutively, independent of IFNa. Furthermore, basic biochemical analysis has found that the protein is highly unstable in a wide variety of storage conditions, and cannot be recovered readily after being dissociated from the cell. Our preliminary studies indicate that Ly6E alters Adenovirus 5 infection of diploid cells. Together these data show that Ly6E is a dynamic protein that can lend insights into Interferon Stimulated Genes and the antiviral response.

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