Filamentation and inhibition of prokaryotic CTP synthase with ligands
Cytidine triphosphate synthase (CTPS) plays a pivotal role in the de novo synthesis of cytidine triphosphate (CTP), a fundamental building block for RNA and DNA that is essential for life. CTPS is capable of directly binding to all four nucleotide triphosphates: adenine triphosphate, uridine triphos...
| Main Authors: | , , |
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| Format: | Journal article |
| Language: | English |
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Wiley Open Access
2024
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| _version_ | 1826313617030512640 |
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| author | Guo, C Wang, Z Liu, J |
| author_facet | Guo, C Wang, Z Liu, J |
| author_sort | Guo, C |
| collection | OXFORD |
| description | Cytidine triphosphate synthase (CTPS) plays a pivotal role in the de novo synthesis of cytidine triphosphate (CTP), a fundamental building block for RNA and DNA that is essential for life. CTPS is capable of directly binding to all four nucleotide triphosphates: adenine triphosphate, uridine triphosphate, CTP, and guanidine triphosphate. Furthermore, CTPS can form cytoophidia in vivo and metabolic filaments in vitro, undergoing regulation at multiple levels. CTPS is considered a potential therapeutic target for combating invasions or infections by viral or prokaryotic pathogens. Utilizing cryo‐electron microscopy, we determined the structure of Escherichia coli CTPS (ecCTPS) filament in complex with CTP, nicotinamide adenine dinucleotide (NADH), and the covalent inhibitor 6‐diazo‐5‐oxo‐ l‐norleucine (DON), achieving a resolution of 2.9 Å. We constructed a phylogenetic tree based on differences in filament‐forming interfaces and designed a variant to validate our hypothesis, providing an evolutionary perspective on CTPS filament formation. Our computational analysis revealed a solvent‐accessible ammonia tunnel upon DON binding. Through comparative structural analysis, we discern a distinct mode of CTP binding of ecCTPS that differs from eukaryotic counterparts. Combining biochemical assays and structural analysis, we determined and validated the synergistic inhibitory effects of CTP with NADH or adenine on CTPS. Our results expand our comprehension of the diverse regulatory aspects of CTPS and lay a foundation for the design of specific inhibitors targeting prokaryotic CTPS. |
| first_indexed | 2024-09-25T04:17:42Z |
| format | Journal article |
| id | oxford-uuid:bfc68a26-b888-495c-9665-01aca6b1a882 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-09-25T04:17:42Z |
| publishDate | 2024 |
| publisher | Wiley Open Access |
| record_format | dspace |
| spelling | oxford-uuid:bfc68a26-b888-495c-9665-01aca6b1a8822024-07-20T15:16:06ZFilamentation and inhibition of prokaryotic CTP synthase with ligandsJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:bfc68a26-b888-495c-9665-01aca6b1a882EnglishJisc Publications RouterWiley Open Access2024Guo, CWang, ZLiu, JCytidine triphosphate synthase (CTPS) plays a pivotal role in the de novo synthesis of cytidine triphosphate (CTP), a fundamental building block for RNA and DNA that is essential for life. CTPS is capable of directly binding to all four nucleotide triphosphates: adenine triphosphate, uridine triphosphate, CTP, and guanidine triphosphate. Furthermore, CTPS can form cytoophidia in vivo and metabolic filaments in vitro, undergoing regulation at multiple levels. CTPS is considered a potential therapeutic target for combating invasions or infections by viral or prokaryotic pathogens. Utilizing cryo‐electron microscopy, we determined the structure of Escherichia coli CTPS (ecCTPS) filament in complex with CTP, nicotinamide adenine dinucleotide (NADH), and the covalent inhibitor 6‐diazo‐5‐oxo‐ l‐norleucine (DON), achieving a resolution of 2.9 Å. We constructed a phylogenetic tree based on differences in filament‐forming interfaces and designed a variant to validate our hypothesis, providing an evolutionary perspective on CTPS filament formation. Our computational analysis revealed a solvent‐accessible ammonia tunnel upon DON binding. Through comparative structural analysis, we discern a distinct mode of CTP binding of ecCTPS that differs from eukaryotic counterparts. Combining biochemical assays and structural analysis, we determined and validated the synergistic inhibitory effects of CTP with NADH or adenine on CTPS. Our results expand our comprehension of the diverse regulatory aspects of CTPS and lay a foundation for the design of specific inhibitors targeting prokaryotic CTPS. |
| spellingShingle | Guo, C Wang, Z Liu, J Filamentation and inhibition of prokaryotic CTP synthase with ligands |
| title | Filamentation and inhibition of prokaryotic CTP synthase with ligands |
| title_full | Filamentation and inhibition of prokaryotic CTP synthase with ligands |
| title_fullStr | Filamentation and inhibition of prokaryotic CTP synthase with ligands |
| title_full_unstemmed | Filamentation and inhibition of prokaryotic CTP synthase with ligands |
| title_short | Filamentation and inhibition of prokaryotic CTP synthase with ligands |
| title_sort | filamentation and inhibition of prokaryotic ctp synthase with ligands |
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