Premium
Evidence for Activation of Host Cell Protein Kinase C Signaling from Proteomics Analysis of Coxiella burnetii‐ Infected THP‐1 Cells
Author(s) -
Funk Cornelius Joel,
Jackson Christa
Publication year - 2018
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.2018.32.1_supplement.866.1
Subject(s) - coxiella burnetii , protein kinase c , biology , protein kinase a , microbiology and biotechnology , marcks , proteomics , signal transduction , kinase , biochemistry , gene
The bacterium Coxiella burnetii can infect cells found in the human lung and cause a respiratory tract infection that manifests as Q‐fever disease. The acute infection typically results in mild flu‐like symptoms that are treatable with antibiotics; however, chronic infections can lead to endocarditis, hepatitis, or death, if left untreated. Bacteria enter alveolar macrophages within a phagosome, but rather than being destroyed by the hydrolytic lysosomal enzymes, C. burnetii induces the host cell to develop a lysosome‐like organelle designated the parasitophorous vacuole (PV) where replication takes place. The PV grows in size to accommodate the increasing number of replicating cells before the bacterial cells are released as the host cell is destroyed. The objective of this study was to identify Protein Kinase C (PKC) substrates and other activated cell signaling pathways using proteomics. Proteins from infected and uninfected THP‐1 cell samples were labeled using isobaric tagging and analyzed using an ion trap mass analyzer (LC‐MS/MS) to identify proteins and post‐translational modifications that were altered during infection. Previous work indicated that C. burnetii infections lead to activation of several Protein Kinase C (PKC) isoforms, which in turn lead to phosphorylation of the PKC substrate MARCKS. Analysis of infected THP‐1 cell samples identified two additional PKC substrates that were phosphorylated during the infection, MARCKSL1 (a MARCKS‐related protein) and ABCA1 (a cholesterol efflux protein). Additional analysis indicated that both proteins were upregulated during the infection at the gene and protein level using qPCR and immunoblotting, respectively. These results provide additional evidence that PKC cell signaling is altered during C. burnetii infection of the host cell and may provide insight into critical events that can be targeted for inhibition of the infection. Support or Funding Information Supported by a grant from Arkansas INBRE (NIH NIGMS P20 GM103429). This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .