We investigated the impact of intracellular hydrogen sulfide (H
2
S) hyperaccumulation on the transcriptome of
Escherichia coli
. The wild-type (WT) strain overexpressing
mstA
, encoding 3-mercaptopyruvate sulfur transferase, produced significantly higher H
2
S levels than the control WT strain. The
mstA
-overexpressing strain exhibited increased resistance to antibiotics, supporting the prior hypothesis that intracellular H
2
S contributes to oxidative stress responses and antibiotic resistance. RNA-seq analysis revealed that over 1,000 genes were significantly upregulated or downregulated upon
mstA
overexpression. The upregulated genes encompassed those associated with iron uptake, including siderophore synthesis and iron import transporters. The
mstA
-overexpressing strain showed increased levels of intracellular iron content, indicating that H
2
S hyperaccumulation affects iron availability within cells. We found that the H
2
S-/supersulfide-responsive transcription factor YgaV is required for the upregulated expression of iron uptake genes in the
mstA
-overexpression conditions. These findings indicate that the expression of iron uptake genes is regulated by intracellular H
2
S, which is crucial for oxidative stress responses and antibiotic resistance in
E. coli
.
IMPORTANCE
H
2
S is recognized as a second messenger in bacteria, playing a vital role in diverse intracellular and extracellular activities, including oxidative stress responses and antibiotic resistance. Both H
2
S and iron serve as essential signaling molecules for gut bacteria. However, the intricate intracellular coordination between them, governing bacterial physiology, remains poorly understood. This study unveils a close relationship between intracellular H
2
S accumulation and iron uptake activity, a relationship critical for antibiotic resistance. We present additional evidence expanding the role of intracellular H2S synthesis in bacterial physiology.
