[PMID]: | 28654654 |
[Au] Autor: | Post DMB; Schilling B; Reinders LM; D'Souza AK; Ketterer MR; Kiel SJ; Chande AT; Apicella MA; Gibson BW |
[Ad] Endereço: | Buck Institute for Research on Aging, Novato, California, United States of America. |
[Ti] Título: | Identification and characterization of AckA-dependent protein acetylation in Neisseria gonorrhoeae. |
[So] Source: | PLoS One;12(6):e0179621, 2017. |
[Is] ISSN: | 1932-6203 |
[Cp] País de publicação: | United States |
[La] Idioma: | eng |
[Ab] Resumo: | Neisseria gonorrhoeae, the causative agent of gonorrhea, has a number of factors known to contribute to pathogenesis; however, a full understanding of these processes and their regulation has proven to be elusive. Post-translational modifications (PTMs) of bacterial proteins are now recognized as one mechanism of protein regulation. In the present study, Western blot analyses, with an anti-acetyl-lysine antibody, indicated that a large number of gonococcal proteins are post-translationally modified. Previous work has shown that Nε-lysine acetylation can occur non-enzymatically with acetyl-phosphate (AcP) as the acetyl donor. In the current study, an acetate kinase mutant (1291ackA), which accumulates AcP, was generated in N. gonorrhoeae. Broth cultures of N. gonorrhoeae 1291wt and 1291ackA were grown, proteins extracted and digested, and peptides containing acetylated-lysines (K-acetyl) were affinity-enriched from both strains. Mass spectrometric analyses of these samples identified a total of 2686 unique acetylation sites. Label-free relative quantitation of the K-acetyl peptides derived from the ackA and wild-type (wt) strains demonstrated that 109 acetylation sites had an ackA/wt ratio>2 and p-values <0.05 in at least 2/3 of the biological replicates and were designated as "AckA-dependent". Regulated K-acetyl sites were found in ribosomal proteins, central metabolism proteins, iron acquisition and regulation proteins, pilus assembly and regulation proteins, and a two-component response regulator. Since AckA is part of a metabolic pathway, comparative growth studies of the ackA mutant and wt strains were performed. The mutant showed a growth defect under aerobic conditions, an inability to grow anaerobically, and a defect in biofilm maturation. In conclusion, the current study identified AckA-dependent acetylation sites in N. gonorrhoeae and determined that these sites are found in a diverse group of proteins. This work lays the foundation for future studies focusing on specific acetylation sites that may have relevance in gonococcal pathogenesis and metabolism. |
[Mh] Termos MeSH primário: |
Acetato Quinase/metabolismo Proteínas de Bactérias/metabolismo Redes e Vias Metabólicas/fisiologia Neisseria gonorrhoeae/metabolismo
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[Mh] Termos MeSH secundário: |
Acetato Quinase/genética Acetilação Proteínas de Bactérias/genética Regulação Bacteriana da Expressão Gênica Espectrometria de Massas Fosforilação Processamento de Proteína Pós-Traducional
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[Pt] Tipo de publicação: | JOURNAL ARTICLE |
[Nm] Nome de substância:
| 0 (Bacterial Proteins); EC 2.7.2.1 (Acetate Kinase) |
[Em] Mês de entrada: | 1709 |
[Cu] Atualização por classe: | 170920 |
[Lr] Data última revisão:
| 170920 |
[Sb] Subgrupo de revista: | IM |
[Da] Data de entrada para processamento: | 170628 |
[St] Status: | MEDLINE |
[do] DOI: | 10.1371/journal.pone.0179621 |
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