Stress granules and RNA processing bodies are novel autoantibody targets in systemic sclerosis

Arthritis Research & Therapy
Michael E JohnsonMichael L Whitfield

Abstract

Autoantibody profiles represent important patient stratification markers in systemic sclerosis (SSc). Here, we performed serum-immunoprecipitations with patient antibodies followed by mass spectrometry (LC-MS/MS) to obtain an unbiased view of all possible autoantibody targets and their associated molecular complexes recognized by SSc. HeLa whole cell lysates were immunoprecipitated (IP) using sera of patients with SSc clinically positive for autoantibodies against RNA polymerase III (RNAP3), topoisomerase 1 (TOP1), and centromere proteins (CENP). IP eluates were then analyzed by LC-MS/MS to identify novel proteins and complexes targeted in SSc. Target proteins were examined using a functional interaction network to identify major macromolecular complexes, with direct targets validated by IP-Western blots and immunofluorescence. A wide range of peptides were detected across patients in each clinical autoantibody group. Each group contained peptides representing a broad spectrum of proteins in large macromolecular complexes, with significant overlap between groups. Network analyses revealed significant enrichment for proteins in RNA processing bodies (PB) and cytosolic stress granules (SG) across all SSc subtypes, which were conf...Continue Reading

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Apr 26, 2016·Arthritis & Rheumatology·Marvin J Fritzler, May Y Choi
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Methods Mentioned

BETA
immunoprecipitations
protein assay
Fusion Tribrid
immunoprecipitation
Profiler
fluorescence microscopy
nucleic
protein folding
from

Software Mentioned

TreeView
: Profiler
Profiler
Cytoscape
gProfiler
Adobe Photoshop
Cluster
igraph
Java
GIANT

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