Successful host defense against pathogens requires innate immune recognition of the correct
pathogen associated molecular patterns (
PAMPs) by pathogen recognition receptors (
PRRs) to trigger the appropriate gene program tailored to the pathogen. While many PRR pathways contribute to the innate immune response to specific pathogens, the relative importance of each pathway for the complete transcriptional program elicited has not been examined in detail. Herein, we used
RNA-sequencing with wildtype and mutant macrophages to delineate the innate immune pathways contributing to the early transcriptional response to Staphylococcus aureus, a ubiquitous microorganism that can activate a wide variety of
PRRs. Unexpectedly, two PRR pathways-the
Toll-like receptor (TLR) and Stimulator of
Interferon Gene (
STING) pathways-were identified as dominant regulators of approximately 95% of the genes that were potently induced within the first four hours of macrophage
infection with live S. aureus. TLR signaling predominantly activated a pro-inflammatory program while
STING signaling activated an
antiviral/
type I interferon response with live but not killed S. aureus. This
STING response was largely dependent on the cytosolic
DNA sensor cyclic
guanosine-
adenosine synthase (cGAS). Using a cutaneous
infection model, we found that the TLR and
STING pathways played opposite roles in host defense to S. aureus. TLR signaling was required for host defense, with its absence reducing
interleukin (IL)-1β production and neutrophil recruitment, resulting in increased bacterial growth. In contrast, absence of
STING signaling had the opposite effect, enhancing the ability to restrict the
infection. These results provide novel insights into the complex interplay of innate immune signaling pathways triggered by S. aureus and uncover opposing roles of TLR and
STING in cutaneous host defense to S. aureus.