Presentation Information
[P03-410]Transcriptome Analysis of Acetophenone-Responsive Genes in Streptomyces coelicolor A3(2)
○SHUMA SUZUKI1, Nanami NAKAJIMA1, Tadashi USUKI1, Taishi TUBOUCHI2, Akito HOSOI3, Shinsaku ITO1, Shunsuke YAJIMA1, Yasuyuki SASAKI1 (1. Tokyo Universityb of Agriculture Faculty of Life Sciences (Japan), 2. Graduate School of Medicine, Osaka Metropolitan University (Japan), 3. NODAI Genome Reserch Center Tokyo Universityb of Agriculture (Japan))
Keywords:
Streptomyces coelicolor A3 (2),MVOC,acetophenone
[Purpose]
Streptomyces coelicolor A3(2) produces acetophenone, a microbial volatile organic compound (MVOC). Exogenous acetophenone increases actinorhodin (ACT) production in S. coelicolor A3(2), suggesting that acetophenone plays a physiological role in this strain. RNA-seq analysis showed that acetophenone treatment broadly altered gene expression and strongly induced sco0230, which encodes a putative transcriptional regulator of unknown function. This study aimed to analyze the transcriptional response to acetophenone and to clarify the function of sco0230.
[Method]
S. coelicolor A3(2) was treated with acetophenone and subjected to RNA-seq and GO enrichment analyses. Among the genes identified by RNA-seq, sco0230 was selected for further analysis, and a sco0230 deletion mutant (Δsco0230) was constructed.ACT production, colony area, and sco0229 expression were compared between the wild type and the mutant. In addition, His-tagged SCO0230 was expressed in Escherichia coli, purified, and analyzed by electrophoretic mobility shift assay (EMSA) using the upstream region of sco0229 as a probe.
[Results]
RNA-seq analysis showed that acetophenone treatment broadly altered gene expression and strongly induced sco0229 and sco0230. GO enrichment analysis identified changes in pathways related to ectoine biosynthesis and metabolism, hydrogen sulfide biosynthesis and metabolism, sulfate metabolism, amino acid and organic acid transport, and signal transduction. Functional analysis of sco0230 showed that the Δsco0230 mutant exhibited increased ACT production and a significantly larger colony area than the wild type. In addition, sco0229 transcription was markedly elevated in the mutant. EMSA demonstrated that SCO0230 directly binds to the upstream region of sco0229.
[Consideration]
The increase in ACT production after acetophenone treatment, together with broad early changes in gene expression, indicates that acetophenone acts upstream of multiple regulatory networks in S. coelicolor A3(2). In particular, changes in pathways related to ectoine biosynthesis and metabolism, sulfur metabolism, and signal transduction support its involvement in stress response, metabolic balance, and secondary metabolism. These findings suggest that acetophenone functions not simply as a metabolic byproduct, but as an endogenous signaling molecule that regulates transcriptional responses and secondary metabolism. Among the acetophenone-responsive genes, sco0230 was strongly induced and is likely to function as a transcriptional repressor of sco0229. SCO0230 is also suggested to be involved in the negative regulation of colony growth and ACT production.
[Conclusion]
Acetophenone may affect multiple regulatory pathways in S. coelicolor A3(2). Among the acetophenone-responsive genes, sco0230 is a candidate regulatory factor that directly represses sco0229 and may be involved in the control of colony growth and ACT production.
Streptomyces coelicolor A3(2) produces acetophenone, a microbial volatile organic compound (MVOC). Exogenous acetophenone increases actinorhodin (ACT) production in S. coelicolor A3(2), suggesting that acetophenone plays a physiological role in this strain. RNA-seq analysis showed that acetophenone treatment broadly altered gene expression and strongly induced sco0230, which encodes a putative transcriptional regulator of unknown function. This study aimed to analyze the transcriptional response to acetophenone and to clarify the function of sco0230.
[Method]
S. coelicolor A3(2) was treated with acetophenone and subjected to RNA-seq and GO enrichment analyses. Among the genes identified by RNA-seq, sco0230 was selected for further analysis, and a sco0230 deletion mutant (Δsco0230) was constructed.ACT production, colony area, and sco0229 expression were compared between the wild type and the mutant. In addition, His-tagged SCO0230 was expressed in Escherichia coli, purified, and analyzed by electrophoretic mobility shift assay (EMSA) using the upstream region of sco0229 as a probe.
[Results]
RNA-seq analysis showed that acetophenone treatment broadly altered gene expression and strongly induced sco0229 and sco0230. GO enrichment analysis identified changes in pathways related to ectoine biosynthesis and metabolism, hydrogen sulfide biosynthesis and metabolism, sulfate metabolism, amino acid and organic acid transport, and signal transduction. Functional analysis of sco0230 showed that the Δsco0230 mutant exhibited increased ACT production and a significantly larger colony area than the wild type. In addition, sco0229 transcription was markedly elevated in the mutant. EMSA demonstrated that SCO0230 directly binds to the upstream region of sco0229.
[Consideration]
The increase in ACT production after acetophenone treatment, together with broad early changes in gene expression, indicates that acetophenone acts upstream of multiple regulatory networks in S. coelicolor A3(2). In particular, changes in pathways related to ectoine biosynthesis and metabolism, sulfur metabolism, and signal transduction support its involvement in stress response, metabolic balance, and secondary metabolism. These findings suggest that acetophenone functions not simply as a metabolic byproduct, but as an endogenous signaling molecule that regulates transcriptional responses and secondary metabolism. Among the acetophenone-responsive genes, sco0230 was strongly induced and is likely to function as a transcriptional repressor of sco0229. SCO0230 is also suggested to be involved in the negative regulation of colony growth and ACT production.
[Conclusion]
Acetophenone may affect multiple regulatory pathways in S. coelicolor A3(2). Among the acetophenone-responsive genes, sco0230 is a candidate regulatory factor that directly represses sco0229 and may be involved in the control of colony growth and ACT production.
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