Proteomic analysis of RAW macrophages treated with cGAMP or c-di-GMP reveals differentially activated cellular pathways
Moloud Aflaki Sooreshjani; Uma K. Aryalc; Ulvi K. Gursoy; Herman O. Sintim
https://urn.fi/URN:NBN:fi-fe2021042720217
Tiivistelmä
Global and quantitative analysis of the proteome help to reveal how host cells sense invading bacteria and
respond to bacterial signaling molecules. Here, we performed label free quantitative proteomic analysis of
RAW macrophages treated with host-derived cGAMP and bacterial-derived c-di-GMP, in an attempt to
identify cellular pathways impacted by these dinucleotides and determine if the host responds
differentially to these two cyclic dinucleotides. We identified a total of 3811 proteins of which
abundances of 404 proteins in cGAMP and 236 proteins in c-di-GMP treated cells were significantly
different compared to the control. Many of the proteins that were strongly and commonly upregulated,
such as interferon-induced proteins 47, 202 and 204 (Ifi47, Ifi202, Ifi204), ubiquitin-activating enzyme E7
(Uba7), interferon-induced protein with tetratricopeptide repeats 1, 2 or 3 (Ifit1, Ifit2, Ifit3), ubiquitin-like
protein ISG15 (ISG15), might be due to the fact that both dinucleotides promote the production of
interferons, which induce the expression of many proteins. However, there were also other proteins that
were differentially affected by cGAMP or c-di-GMP treatment, including probable ATP-dependent RNA
helicase DHX58 (Dhx58), nuclear autoantigen Sp-100 (Sp100), MARCKS-related protein (Marcksl1) and
antigen peptide transporter 2 (Tap2). This is probably due to the differential levels of IFNs produced by
the dinucleotides or may indicate that non-STING activation might also contribute to the host's response
to c-di-GMP and cGAMP. Interestingly Trex1, a nuclease that degrades DNA (an activator of cGAS to
produce cGAMP), was upregulated (3.22 fold) upon cGAMP treatment, hinting at a possible feedback
loop to regulate cGAMP synthesis. These results lay a foundation for future studies to better characterize
and understand the complex c-di-GMP and cGAMP signaling network.
Kokoelmat
- Rinnakkaistallenteet [19207]