The Fic protein Doc uses an inverted substrate to phosphorylate and inactivate EF-Tu

Castro-Roa, Daniel; García-Pino, Abel; Gieter, Steven De; Nuland, Nico A. J. van; Loris, Remy; Zenkin, Nikolay

doi:10.1038/nchembio.1364

Cited by 159 publications

(187 citation statements)

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“…AvrAC from the plant pathogen Xanthomonas campestris covalently links UMP to RIPK and BIK1 kinases, thereby interfering with host immune defense (13). Finally, the bacterial Fic protein Doc, the toxin unit of the Doc-PhD toxin-antitoxin system present in E. coli, phosphorylates Thr-382 of the translation elongation factor EF-Tu, inhibiting translation (14,15). Thus, Fic protein activity is not linked exclusively to the transfer of AMP onto target proteins.…”

Section: All Heavy Chain-only Antibody Variable Domains Bind Hype Whementioning

confidence: 99%

HypE-specific Nanobodies as Tools to Modulate HypE-mediated Target AMPylation

Truttmann

Qin

Stiegeler

et al. 2015

Journal of Biological Chemistry

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Section: All Heavy Chain-only Antibody Variable Domains Bind Hype Whementioning

confidence: 99%

HypE-specific Nanobodies as Tools to Modulate HypE-mediated Target AMPylation

Truttmann

Qin

Stiegeler

et al. 2015

Journal of Biological Chemistry

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“…Antitoxins are proteolytically degraded after (p)ppGpp accumulation, leading to derepression at the toxin-antitoxin promoter (8,12). Liberated toxin proteins inhibit the replication or translation machinery by targeting DNA gyrase, initiator tRNA fMet , glutamyl-tRNA synthetase, EF-Tu, free mRNA, ribosome-bound mRNA, and the ribosome itself (13)(14)(15)(16)(17)(18)(19)(20).…”

mentioning

confidence: 99%

Defining the mRNA recognition signature of a bacterial toxin protein

Schureck

Dunkle

Maehigashi

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Bacteria contain multiple type II toxins that selectively degrade mRNAs bound to the ribosome to regulate translation and growth and facilitate survival during the stringent response. Ribosomedependent toxins recognize a variety of three-nucleotide codons within the aminoacyl (A) site, but how these endonucleases achieve substrate specificity remains poorly understood. Here, we identify the critical features for how the host inhibition of growth B (HigB) toxin recognizes each of the three A-site nucleotides for cleavage. X-ray crystal structures of HigB bound to two different codons on the ribosome illustrate how HigB uses a microbial RNase-like nucleotide recognition loop to recognize either cytosine or adenosine at the second A-site position. Strikingly, a single HigB residue and 16S rRNA residue C1054 form an adenosine-specific pocket at the third A-site nucleotide, in contrast to how tRNAs decode mRNA. Our results demonstrate that the most important determinant for mRNA cleavage by ribosome-dependent toxins is interaction with the third A-site nucleotide. (1,3,4). This rapid inhibitory switch suppresses metabolite consumption and temporarily halts cell growth to promote bacterial survival until nutrients are readily available. Among the prosurvival genes regulated by (p)ppGpp production are toxin-antitoxin modules, which have additional roles in antibiotic resistance and tolerance, biofilm and persister cell formation, and niche-specific colonization (5-11). The critical roles toxin-antitoxin pairs play in bacterial physiology underscore the importance of understanding their molecular targets and modes of action.There are five different classes (I to V) of toxin-antitoxin systems defined by how the antitoxin represses toxin function (1). Type II toxin-antitoxin pairs form protein-protein complexes during exponential growth that serve two purposes: inhibition of toxin activity by antitoxin binding and transcriptional autorepression to limit toxin expression (12). Antitoxins are proteolytically degraded after (p)ppGpp accumulation, leading to derepression at the toxin-antitoxin promoter (8, 12). Liberated toxin proteins inhibit the replication or translation machinery by targeting DNA gyrase, initiator tRNA fMet , glutamyl-tRNA synthetase, EF-Tu, free mRNA, ribosome-bound mRNA, and the ribosome itself (13-20).Ribosome-dependent toxins cleave mRNAs on the ribosome between the second and third nucleotides of the aminoacyl (A)-site codon (21-23). Although collectively Escherichia coli ribosome-dependent toxins target a diverse range of codons, each individual toxin appears to have a strong codon preference and cleaves at defined positions along the mRNA (24-26). RelE cleaves at UAG stop codons and the CAG sense codon (all codons denoted in the 5′-3′ direction); YoeB cleaves at codons following a translational AUG start site and at the UAA stop codon; and YafQ cleaves a single AAA sense codon (16,24,(27)(28)(29). In contrast, Proteus vulgaris host inhibition of growth B (HigB) toxin degrades multiple codons encod...

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“…AvrAC from the plant pathogen Xanthomonas campestris adds UMP to the host kinases BIK1 and RIPK to suppress the host immune response (10). The bacteriophage toxin Doc, which belongs to a distant subfamily of Fic proteins, is a kinase that inhibits bacterial translation by phosphorylating the translation elongation factor EF-Tu (11,12). Structural analysis has shown that the versatility of the Fic domain for AMPylation, UMPylation, phosphorylation, and phosphocholination occurs by changing the orientation of the nucleotide-based substrates in the active site (13).…”

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confidence: 99%