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. Author manuscript; available in PMC: 2019 May 9.
Published in final edited form as: Cell Host Microbe. 2018 May 9;23(5):594–606.e7. doi: 10.1016/j.chom.2018.04.001

Figure 4. S-nitrosylation of AgrA Disrupts Agr Auto-Activation by Decreasing AgrA Occupancy at Target Promoters.

Figure 4

(A) Western blot of HA-tagged AgrA from S. aureus cultures treated with NO (n=3).

(B) Western blot of HA-tagged AgrA from S. aureus cultures treated with NO after inhibition of protein synthesis with tetracycline (n=2).

(C) RT-qPCR of agrA and RNAIII from cultures ectopically expressing AgrA and activated with spent medium preceding NO treatment (n=3).

(D) ChIP-qPCR of NO treated S. aureus wild-type and agrA C55S C199S cultures (n=3). DNA regions proximal to AgrA binding sites include agrIR and RNAIII; control regions include rpoD, gmk, and gyrB. All chromosomal regions are normalized to the ChIPc control region located several kilobases away from agr intergenic region. The numbers above bars indicate fold difference between the means of indicated samples. The p values comparing the NO-dependent differences (untreated vs. NO-treated) between WT and agrA C55S C199S are 0.045 for agrIR and 0.25 for RNAIII.

Student’s t test, *p<0.05; ns, not significant.

For (A) through (D) data are represented as means with error bars showing standard deviation..

See also Figure S3.