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Determining Cellular and Viral Interaction Partners of the Kaposi's Sarcoma‐associated Herpesvirus Protein ORF55
Author(s) -
Randolph Keyera,
Rivas Hembly,
Gaglia Marta Maria
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.600.13
Subject(s) - kaposi's sarcoma associated herpesvirus , biology , gene , regulator , immunoprecipitation , reporter gene , transcriptional regulation , gene expression , regulation of gene expression , viral protein , microbiology and biotechnology , virology , virus , genetics , herpesviridae , viral disease
Kaposi's sarcoma (KS) is a cancer caused by Kaposi's sarcoma‐associated herpesvirus (KSHV). KS mainly affects immunocompromised patients such as AIDS patients and transplant patients. There are few available treatments for KS and none of them directly targets KSHV infection. KSHV alters host gene expression, a process thought to promote viral replication. At present the viral factors involved in the regulation of host gene expression are unknown. Our lab screened for viral factors that are involved in host gene modulation using reporter assays and identified the KSHV protein ORF55 as a potential new gene regulator. We found that ORF55 causes reporter RNA and protein levels to decrease by two‐fold. We hypothesize that ORF55 plays a role in gene regulation through interactions with cellular proteins. We used the results of a published mass‐spectrometry screen of cellular proteins that interact with 90 different KSHV proteins to select candidate ORF55‐interacting proteins, and we focused on several proteins involved in signaling proteins and transcriptional regulation. We overexpressed ORF55 and tagged version of the candidate interacting cellular proteins and used co‐immunoprecipitation followed by western blotting to determine if these proteins are interacting with ORF55. Our results suggest that ORF55 does not interact with RACK1, 14‐3‐3 protein ɛ and the transcriptional regulator TRIM28. Further studies will be performed for the remaining cellular proteins of interest.