Common Mechanism for Target Specificity of Protein- and DNA-Targeting ADP-Ribosyltransferases

Yoshida, Toru; Tsuge, Hideaki

doi:10.3390/toxins13010040

Cited by 10 publications

(7 citation statements)

References 65 publications

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“…In other members of the R-S-E clade of bacterial ARTs 24 , the residue (ϕ) in the third position in the ADP-ribosylating toxin turn-turn (ARTT) loop motif X-X-ϕ-X-X-E/Q-X-E, forms essential hydrophobic interactions with substrates. Although the corresponding residue (P260) of TcART, sits at the interface to F-actin, it does not interact there with any specific residue or with a hydrophobic patch (Fig 2c, S7f).…”

Section: Resultsmentioning

confidence: 99%

Mechanism of threonine ADP-ribosylation of F-actin by a Tc toxin

Belyy

Lindemann

Roderer

et al. 2022

Preprint

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Tc toxins deliver toxic enzymes into host cells by a unique injection mechanism. One of these enzymes is TccC3, an ADP-ribosyltransferase from Photorhabdus luminescens. Once TccC3 is translocated into the target cell, the enzyme ADP-ribosylates actin, resulting in clustering of the actin cytoskeleton and ultimately cell death. Here, we combine biochemistry, solution and solid-state NMR spectroscopy and cryo-EM to show in atomic detail how TccC3 modifies actin. We find that the ADP-ribosyltransferase does not bind to G-actin but interacts with two consecutive actin subunits of F-actin. The binding of TccC3 to F-actin occurs via an induced-fit mechanism that facilitates access of NAD+ to the nucleotide binding pocket. The following nucleophilic substitution reaction results in the transfer of ADP-ribose to threonine-148 of F-actin. We demonstrate that this site-specific modification of F-actin prevents its interaction with depolymerization factors, such as cofilin, which impairs actin network turnover and leads to steady actin polymerization. Our findings reveal in atomic detail a new mechanism of action of a bacterial toxin through specific targeting and modification of F-actin.

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Section: Resultsmentioning

confidence: 99%

Mechanism of threonine ADP-ribosylation of F-actin by a Tc toxin

Belyy

Lindemann

Roderer

et al. 2022

Preprint

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“…The glutamic acid residue of this motif, which is a critical catalytic residue in other ARTs, such as diphtheria toxin and PARP-1 13 , is replaced in Arr by an aspartate residue (D84 in Fig. 3 ) 1 , 14 . Here, we observed that this H-Y-[QED] motif is highly conserved in almost all hundreds of Arr sequences analyzed, indicating its importance in Arr activity, and suggesting that these Arr sequences could be functional.…”

Section: Discussionmentioning

confidence: 99%

Genomic epidemiology of rifampicin ADP-ribosyltransferase (Arr) in the Bacteria domain

Morgado

Fonseca

Vicente

2021

Sci Rep

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Arr is an ADP-ribosyltransferase enzyme primarily reported in association with rifamycin resistance, which has been used to treat tuberculosis in addition to Gram-positive infections and, recently, pan-resistant Gram-negative bacteria. The arr gene was initially identified on the Mycolicibacterium smegmatis chromosome and later on Proteobacteria plasmids. This scenario raised concerns on the distribution and spread of arr, considering the Bacteria domain. Based on 198,082 bacterial genomes/metagenomes, we performed in silico analysis, including phylogenetic reconstruction of Arr in different genomic contexts. Besides, new arr alleles were evaluated by in vitro analysis to assess their association with rifampin resistance phenotype. The arr gene was prevalent in thousands of chromosomes and in hundreds of plasmids from environmental and clinical bacteria, mainly from the phyla Actinobacteria, Proteobacteria, Firmicutes, and Bacteroidetes. Furthermore, this gene was identified in other and new genomic contexts. Interestingly, Arr sequences associated with rifampin resistance were distributed across all phylogeny, indicating that, despite the diversity, their association with rifampin resistance phenotype were maintained. In fact, we found that the key residues were highly conserved. In addition, other analyzes have raised evidence of another Arr function, which is related to guanidine metabolism. Finally, this scenario as a whole also suggested the Actinobacteria phylum as a potential ancestral source of arr within the Bacteria domain.

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“…失 [22,23] . 在整个反应过程中, NAD + 被固定在核心区域形成的疏水裂隙结合口袋中 [24] . 此外, 还有一类在结构和催化基序上不同, 但依赖NAD + 的脱乙酰化酶-Sirtuin家族, 也具有ART活性, 及一些非经典类型的…”

Section: 细菌Arts的结构特点unclassified