A dual program for CRP-mediated regulation in bacterial alarmone (p)ppGpp
ABSTRACT Gene expression and proper downstream cellular functions upon facing environmental shifts depend on the combined and cooperative regulation of genetic networks. Here, we identified cAMP receptor protein (CRP) as a master regulator of (p)ppGpp (guanosine tetra- and penta-phosphate) homeostas...
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American Society for Microbiology
2024-11-01
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| Series: | mBio |
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| Online Access: | https://journals.asm.org/doi/10.1128/mbio.02430-24 |
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| author | Li Zhao Shi-Yu Zhou Yu Fu Jin-Long Shen Bin-Cheng Yin Di You Bang-Ce Ye |
| author_facet | Li Zhao Shi-Yu Zhou Yu Fu Jin-Long Shen Bin-Cheng Yin Di You Bang-Ce Ye |
| author_sort | Li Zhao |
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| description | ABSTRACT Gene expression and proper downstream cellular functions upon facing environmental shifts depend on the combined and cooperative regulation of genetic networks. Here, we identified cAMP receptor protein (CRP) as a master regulator of (p)ppGpp (guanosine tetra- and penta-phosphate) homeostasis. Via CRP-mediated direct transcriptional regulation of the (p)ppGpp synthetase/hydrolase RelA and SpoT, cAMP-CRP stimulates pervasive accumulation of (p)ppGpp under glucose-limiting conditions. Notably, CRP exerts a nonclassical property as a translational regulator through YfiQ-dependent acetylation of ribosome protein S1 at K247, which further enhances the translation of RelA, SpoT, and CRP itself. From a synthetic biology perspective, this self-activating feedback loop for (p)ppGpp synthesis highlights the function of CRP-mediated dual enhancement (CMDE) in controlling bacterial gene expression, which enables stable activation of genetic circuits. CMDE applied in synthetic circuits leads to a stable increase in p-coumaric acid, cinnamic acid, and pinosylvin production. Our findings showed that CRP-mediated dual circuits for (p)ppGpp regulation enable robust activation that could address bioproduction and other biotechnological needs.IMPORTANCETranscriptional–translational coordination is fundamental for rapid and efficient gene expression in most bacteria. Here, we uncovered the roles of cAMP-CRP in this process. We found that CRP distinctly increases RelA and SpoT transcription and translation, and that acetylation of S1 at K247 accelerates the self-activation of the leading CRP under glucose-limiting conditions. We further found that elevated (p)ppGpp significantly impedes the formation of the cAMP-CRP complex, an active form responsible for transcriptional activation. A model was created in which cAMP-CRP and (p)ppGpp cooperate to dynamically modulate the efficiency of transcriptional–translational coordination responses to stress. More broadly, productive activation in synthetic circuits was achieved through the application of CRP-mediated dual enhancement (CMDE), promising to inspire new approaches for the development of cell-based biotechnologies. |
| format | Article |
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| institution | OA Journals |
| issn | 2150-7511 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | American Society for Microbiology |
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| series | mBio |
| spelling | doaj-art-0edb2375aee44b24999e8039a3cc0de22025-08-20T02:14:38ZengAmerican Society for MicrobiologymBio2150-75112024-11-01151110.1128/mbio.02430-24A dual program for CRP-mediated regulation in bacterial alarmone (p)ppGppLi Zhao0Shi-Yu Zhou1Yu Fu2Jin-Long Shen3Bin-Cheng Yin4Di You5Bang-Ce Ye6Lab of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, ChinaLab of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, ChinaLab of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, ChinaLab of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, ChinaLab of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, ChinaLab of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, ChinaLab of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, ChinaABSTRACT Gene expression and proper downstream cellular functions upon facing environmental shifts depend on the combined and cooperative regulation of genetic networks. Here, we identified cAMP receptor protein (CRP) as a master regulator of (p)ppGpp (guanosine tetra- and penta-phosphate) homeostasis. Via CRP-mediated direct transcriptional regulation of the (p)ppGpp synthetase/hydrolase RelA and SpoT, cAMP-CRP stimulates pervasive accumulation of (p)ppGpp under glucose-limiting conditions. Notably, CRP exerts a nonclassical property as a translational regulator through YfiQ-dependent acetylation of ribosome protein S1 at K247, which further enhances the translation of RelA, SpoT, and CRP itself. From a synthetic biology perspective, this self-activating feedback loop for (p)ppGpp synthesis highlights the function of CRP-mediated dual enhancement (CMDE) in controlling bacterial gene expression, which enables stable activation of genetic circuits. CMDE applied in synthetic circuits leads to a stable increase in p-coumaric acid, cinnamic acid, and pinosylvin production. Our findings showed that CRP-mediated dual circuits for (p)ppGpp regulation enable robust activation that could address bioproduction and other biotechnological needs.IMPORTANCETranscriptional–translational coordination is fundamental for rapid and efficient gene expression in most bacteria. Here, we uncovered the roles of cAMP-CRP in this process. We found that CRP distinctly increases RelA and SpoT transcription and translation, and that acetylation of S1 at K247 accelerates the self-activation of the leading CRP under glucose-limiting conditions. We further found that elevated (p)ppGpp significantly impedes the formation of the cAMP-CRP complex, an active form responsible for transcriptional activation. A model was created in which cAMP-CRP and (p)ppGpp cooperate to dynamically modulate the efficiency of transcriptional–translational coordination responses to stress. More broadly, productive activation in synthetic circuits was achieved through the application of CRP-mediated dual enhancement (CMDE), promising to inspire new approaches for the development of cell-based biotechnologies.https://journals.asm.org/doi/10.1128/mbio.02430-24cAMP-CRP(p)ppGppprotein acetylationtranscriptional regulationtranslational regulation |
| spellingShingle | Li Zhao Shi-Yu Zhou Yu Fu Jin-Long Shen Bin-Cheng Yin Di You Bang-Ce Ye A dual program for CRP-mediated regulation in bacterial alarmone (p)ppGpp mBio cAMP-CRP (p)ppGpp protein acetylation transcriptional regulation translational regulation |
| title | A dual program for CRP-mediated regulation in bacterial alarmone (p)ppGpp |
| title_full | A dual program for CRP-mediated regulation in bacterial alarmone (p)ppGpp |
| title_fullStr | A dual program for CRP-mediated regulation in bacterial alarmone (p)ppGpp |
| title_full_unstemmed | A dual program for CRP-mediated regulation in bacterial alarmone (p)ppGpp |
| title_short | A dual program for CRP-mediated regulation in bacterial alarmone (p)ppGpp |
| title_sort | dual program for crp mediated regulation in bacterial alarmone p ppgpp |
| topic | cAMP-CRP (p)ppGpp protein acetylation transcriptional regulation translational regulation |
| url | https://journals.asm.org/doi/10.1128/mbio.02430-24 |
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