Reiterative synthesis by the ribosome and recognition of the N-terminal formyl group by biosynthetic machinery contribute to evolutionary conservation of the length of antibiotic microcin C peptide precursor
Microcin C (McC) is a peptide adenylate antibiotic produced by Escherichia coli cells bearing a plasmid-borne mcc gene cluster. Most MccA precursors, encoded by validated mcc operons from diverse bacteria, are 7 amino acids long, but the significance of this precursor length conservation has remaine...
| Main Authors: | , , , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
American Society for Microbiology
2019
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| _version_ | 1826289360766500864 |
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| author | Zukher, I Pavlov, M Tsibulskaya, D Kulikovsky, A Zyubko, T Bikmetov, D Serebryakova, M Nair, S Ehrenberg, M Dubiley, S Severinov, K |
| author_facet | Zukher, I Pavlov, M Tsibulskaya, D Kulikovsky, A Zyubko, T Bikmetov, D Serebryakova, M Nair, S Ehrenberg, M Dubiley, S Severinov, K |
| author_sort | Zukher, I |
| collection | OXFORD |
| description | Microcin C (McC) is a peptide adenylate antibiotic produced by Escherichia coli cells bearing a plasmid-borne mcc gene cluster. Most MccA precursors, encoded by validated mcc operons from diverse bacteria, are 7 amino acids long, but the significance of this precursor length conservation has remained unclear. Here, we created derivatives of E. coli mcc operons encoding longer precursors and studied their synthesis and bioactivities. We found that increasing the precursor length to 11 amino acids and beyond strongly decreased antibiotic production. We found this decrease to depend on several parameters. First, reiterative synthesis of the MccA peptide by the ribosome was decreased at longer mccA open reading frames, leading to less efficient competition with other messenger RNAs. Second, the presence of a formyl group at the N-terminal methionine of the heptameric peptide had a strong stimulatory effect on adenylation by the MccB enzyme. No such formyl group stimulation was observed for longer peptides. Finally, the presence of the N-terminal formyl on the heptapeptide adenylate stimulated bioactivity, most likely at the uptake stage. Together, these factors should contribute to optimal activity of McC-like compounds as 7-amino-acid peptide moieties and suggest convergent evolution of several steps of the antibiotic biosynthesis pathway and their adjustment to sensitive cell uptake machinery to create a potent drug. |
| first_indexed | 2024-03-07T02:27:42Z |
| format | Journal article |
| id | oxford-uuid:a6310811-6455-4312-a5be-bde4692c4774 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T02:27:42Z |
| publishDate | 2019 |
| publisher | American Society for Microbiology |
| record_format | dspace |
| spelling | oxford-uuid:a6310811-6455-4312-a5be-bde4692c47742022-03-27T02:45:27ZReiterative synthesis by the ribosome and recognition of the N-terminal formyl group by biosynthetic machinery contribute to evolutionary conservation of the length of antibiotic microcin C peptide precursorJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:a6310811-6455-4312-a5be-bde4692c4774EnglishSymplectic Elements at OxfordAmerican Society for Microbiology2019Zukher, IPavlov, MTsibulskaya, DKulikovsky, AZyubko, TBikmetov, DSerebryakova, MNair, SEhrenberg, MDubiley, SSeverinov, KMicrocin C (McC) is a peptide adenylate antibiotic produced by Escherichia coli cells bearing a plasmid-borne mcc gene cluster. Most MccA precursors, encoded by validated mcc operons from diverse bacteria, are 7 amino acids long, but the significance of this precursor length conservation has remained unclear. Here, we created derivatives of E. coli mcc operons encoding longer precursors and studied their synthesis and bioactivities. We found that increasing the precursor length to 11 amino acids and beyond strongly decreased antibiotic production. We found this decrease to depend on several parameters. First, reiterative synthesis of the MccA peptide by the ribosome was decreased at longer mccA open reading frames, leading to less efficient competition with other messenger RNAs. Second, the presence of a formyl group at the N-terminal methionine of the heptameric peptide had a strong stimulatory effect on adenylation by the MccB enzyme. No such formyl group stimulation was observed for longer peptides. Finally, the presence of the N-terminal formyl on the heptapeptide adenylate stimulated bioactivity, most likely at the uptake stage. Together, these factors should contribute to optimal activity of McC-like compounds as 7-amino-acid peptide moieties and suggest convergent evolution of several steps of the antibiotic biosynthesis pathway and their adjustment to sensitive cell uptake machinery to create a potent drug. |
| spellingShingle | Zukher, I Pavlov, M Tsibulskaya, D Kulikovsky, A Zyubko, T Bikmetov, D Serebryakova, M Nair, S Ehrenberg, M Dubiley, S Severinov, K Reiterative synthesis by the ribosome and recognition of the N-terminal formyl group by biosynthetic machinery contribute to evolutionary conservation of the length of antibiotic microcin C peptide precursor |
| title | Reiterative synthesis by the ribosome and recognition of the N-terminal formyl group by biosynthetic machinery contribute to evolutionary conservation of the length of antibiotic microcin C peptide precursor |
| title_full | Reiterative synthesis by the ribosome and recognition of the N-terminal formyl group by biosynthetic machinery contribute to evolutionary conservation of the length of antibiotic microcin C peptide precursor |
| title_fullStr | Reiterative synthesis by the ribosome and recognition of the N-terminal formyl group by biosynthetic machinery contribute to evolutionary conservation of the length of antibiotic microcin C peptide precursor |
| title_full_unstemmed | Reiterative synthesis by the ribosome and recognition of the N-terminal formyl group by biosynthetic machinery contribute to evolutionary conservation of the length of antibiotic microcin C peptide precursor |
| title_short | Reiterative synthesis by the ribosome and recognition of the N-terminal formyl group by biosynthetic machinery contribute to evolutionary conservation of the length of antibiotic microcin C peptide precursor |
| title_sort | reiterative synthesis by the ribosome and recognition of the n terminal formyl group by biosynthetic machinery contribute to evolutionary conservation of the length of antibiotic microcin c peptide precursor |
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