Structure and mechanism of TagA, a novel membrane-associated glycosyltransferase that produces wall teichoic acids in pathogenic bacteria

Kattke, Michele D.; Gosschalk, Jason E.; Martinez, Orlando E.; Kumar, Garima; Gale, Robert T.; Cascio, Duilio; Sawaya, M.R.; Phillips, Martin; Brown, Eric D.; Clubb, Robert T.

doi:10.1101/507483

Cited by 5 publications

(41 citation statements)

References 35 publications

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“…Most pronounced of all functions were cell wall modification associated genes. Within the top five scored genes, we found Mur ligase family [34] and bacitracin resistance [35] proteins associated with resistant strains, and glycosyl transferase family [36] and O-mannosyltrasferase [37] proteins from susceptible strains ( Table 1). These genes may directly or indirectly influence the TM7x attachment to the host.…”

Section: Comparative Genomics Reveal Functional Characteristics Of DImentioning

confidence: 99%

The saccharibacterium TM7x elicits differential responses across its host range

Utter

Cavanaugh

et al. 2020

The ISME Journal

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Host range is a fundamental component of symbiotic interactions, yet it remains poorly characterized for the prevalent yet enigmatic subcategory of bacteria/bacteria symbioses. The recently characterized obligate bacterial epibiont Candidatus Nanosynbacter lyticus TM7x with its bacterial host Actinomyces odontolyticus XH001 offers an ideal system to study such a novel relationship. In this study, the host range of TM7x was investigated by coculturing TM7x with various related Actinomyces strains and characterizing their growth dynamics from initial infection through subsequent co-passages. Of the twenty-seven tested Actinomyces, thirteen strains, including XH001, could host TM7x, and further classified into “permissive” and “nonpermissive” based on their varying initial responses to TM7x. Ten permissive strains exhibited growth/crash/recovery phases following TM7x infection, with crash timing and extent dependent on initial TM7x dosage. Meanwhile, three nonpermissive strains hosted TM7x without a growth-crash phase despite high TM7x dosage. The physical association of TM7x with all hosts, including nonpermissive strains, was confirmed by microscopy. Comparative genomic analyses revealed distinguishing genomic features between permissive and nonpermissive hosts. Our results expand the concept of host range beyond a binary to a wider spectrum, and the varying susceptibility of Actinomyces strains to TM7x underscores how small genetic differences between hosts can underly divergent selective trajectories.

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Section: Comparative Genomics Reveal Functional Characteristics Of DImentioning

confidence: 99%

The saccharibacterium TM7x elicits differential responses across its host range

Utter

Cavanaugh

et al. 2020

The ISME Journal

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“…In previously reported crystal structures, T. italicus TagA ΔC (TagA, residues M1-G195) adopted both dimeric and trimeric oligomeric states ( 36 ). To probe its oligomerization state in solution, we used NMR.…”

Section: Resultsmentioning

confidence: 99%

“…Members of this large family are classified as GT26 enzymes in the Carbohydrate-Active enZYmes database ( www.cazy.org ), but little is known about the molecular basis of their enzymatic activity ( 32 , 33 , 34 , 35 ). Recently, we reported the structure of the TagA enzyme from Thermoanaerobacter italicus , which represents the only known structure of a GT26 enzyme ( 36 , 37 ). The GT26 domain adopts a unique fold that is distinct from other GTs ( e.g.…”

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confidence: 99%

“…, GT-A, GT-B, GT-C, and GT-D folds) and is followed by a 49-amino acid C-terminal tail (CTT) that targets the enzyme to the cell membrane ( 36 ). Functional studies showed that the CTT is essential for activity and identified D65 and R221 as important determinants for catalysis as their alteration causes more than a 20-fold reduction in activity as compared with the wildtype enzyme ( 36 ). However, the molecular basis of substrate recognition and catalysis remains incompletely understood.…”

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Insight into the molecular basis of substrate recognition by the wall teichoic acid glycosyltransferase TagA

Martinez

Mahoney

Goring

et al. 2022

Journal of Biological Chemistry

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This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

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“…Esta foi então purificada por afinidade, com concentrações de imidazol para eluição de 500 SA16 possui 231 aminoácidos, tendo seu peso molecular calculado pelo programa ProtParam 91 de aproximadamente 26 kDa e pI teóricode 9,55 (Figura 43).Além disso, a fluorescência pode ser devida à presença da proteína de carreadora tiorredoxina, que possui 2 triptofanos em sua estrutura primária.Como não foi possível obter a enzima TarA de S. aureus solúvel sem a adição de detergentes e visto que a partir dos experimentos realizados não é possível afirmar que a mesma se encontra enovelada em solução, não foram feitos mais testes com a mesma. Além disso, em 2019, Kattke et al92 publicaram a resolução da estrutura cristalográfica da proteína TagA de Thermoanaerobacter italicus, a qual possui cerca de 37% de identidade com a enzima de S. aureus, incluindo o domínio WecB/TagA/CpsF. Kattke et al Pseudomonas aeruginosa.que possuem enzimas similares àquela de S. aureus.…”

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UDP-N-acetilglicosamina 2-epimerase de >i<Staphylococcus aureus>/i<: estrutura, dinâmica e prospecção de novos ligantes

Azevedo¹

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Staphylococcus aureus (a Gram-positive bacteria) is a major cause of antibiotic-resistant infections in healthcare systems worldwide. It has been shown that the wall teichoic acid (WTA) presents an important target for the development of drugs that fight resistant infections, especially beta-lactam antibiotics. An UDP-N-acetylglycosamine (UDP-GlcNac) 2-epimerase enzyme (or MnaA) from S. aureus, is one of the first enzymes of the WTA biosynthesis pathway. In this work, simulations of molecular dynamics were performed using the crystallographic structure of the enzyme to characterize conformational changes as they occur due to the presence or absence of the UDP (cofactor) and the UDP-GlcNac (substrate). Using different simulation techniques, such as accelerated molecular dynamics by adding a biased potential (ABMD) and adding Gaussian potentials to the total or dihedral potential of the system (GAMD), it was possible to access the energy profile of the systems with and without ligands. The data collected in more than 32 µs of simulation allowed us to analyze the role of the positively charged amino acids present in the active in the movement between the domains and in the maintenance of the substrate in the catalytic site. Also, the protein has an active energy profile which is modified by the presence of UDP / UDP-GlcNac molecules and could lead to understanding the mechanisms in which the substrate enters and/or exits the active site. A search for ligands and possible inhibitors was carried out using the LiBELA docking software, where 17 possible ligands were found. In vitro tests were made using the recombinant protein in differential scanning fluorimetry (DSF) and quantitative DSF (qDSF) assays. An interaction between MnaA and the beta-lactam antibiotic, ticarcillin, was found, leading to the possibility of a new pharmacodynamics associated with this drug, that is already commercially available.

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Structure and mechanism of TagA, a novel membrane-associated glycosyltransferase that produces wall teichoic acids in pathogenic bacteria

Cited by 5 publications

References 35 publications

The saccharibacterium TM7x elicits differential responses across its host range

The saccharibacterium TM7x elicits differential responses across its host range

Insight into the molecular basis of substrate recognition by the wall teichoic acid glycosyltransferase TagA

UDP-N-acetilglicosamina 2-epimerase de >i<Staphylococcus aureus>/i<: estrutura, dinâmica e prospecção de novos ligantes

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