Regulation of Translation of the Head Protein of T4 Bacteriophage by Specific Binding of EF-Tu to a Leader Sequence

Snyder, Larry; Blight, Sherry Kathleen; Auchtung, Jennifer M.

doi:10.1016/j.jmb.2003.09.063

Cited by 10 publications

(11 citation statements)

References 30 publications

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“…Bingham et al (25) have postulated that complexation between EF-Tu and Gol may be required for the maturation of the phage capsid. This is supported by recent work (36) in which EF-Tu was found to associate with the Gol region of gp23 during protein synthesis, its cleavage by Lit blocking the translation of downstream reporters. Snyder et al (36) proposed a model whereby the binding of EF-Tu to the Gol sequence arrests translation and allows the remainder of the gp23 polypeptide to bind the GroEL chaperone, which is required for capsid assembly before translation continues.…”

Section: Discussionsupporting

confidence: 71%

“…This is supported by recent work (36) in which EF-Tu was found to associate with the Gol region of gp23 during protein synthesis, its cleavage by Lit blocking the translation of downstream reporters. Snyder et al (36) proposed a model whereby the binding of EF-Tu to the Gol sequence arrests translation and allows the remainder of the gp23 polypeptide to bind the GroEL chaperone, which is required for capsid assembly before translation continues. This model has some similarities with the translational arrest mechanisms observed for the human immunodeficiency virus and the herpes simplex virus and points to a more general association between viral proteins and elongation translation factors that appears to be required for virus maturation.…”

Section: Discussionsupporting

confidence: 71%

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The Role of an Activating Peptide in Protease-mediated Suicide of Escherichia coli K12

Copeland

Kleanthous

2005

Journal of Biological Chemistry

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Activation of latent proteinases ensures that the timing of proteolysis is regulated precisely, a process that generally involves proteolytic excision of a pro-region or a tightly bound inhibitor. Here we define the activation mechanism for Lit, a dormant suicide proteinase in Escherichia coli K-12. Previous work has shown that Gol, a short sequence within the major capsid protein gp23, activates Lit during the latter stages of T4 phage infection. This results in cell death and exclusion of the phage from the culture. The Lit site specifically cleaves the host translation factor EF-Tu (elongation factor Tu) after it has formed a weak complex with Gol, which can be supplied as a 29-residue peptide. Gol is absolutely required for Lit activation. but its role in proteolysis is unknown. Using a purified three-component system and kinetic analysis, we demonstrate that under physiological conditions Lit hydrolyzes its substrate very slowly (k cat of ϳ1 s ؊1 ). Given the abundance of EF-Tu in the cell, this finding is consistent with a cell-killing mechanism in which a few cleaved EF-Tu proteins are able block translating ribosomes from functioning. We also demonstrate that less than half of the 29 Gol residues are needed for Lit activation and that the role of the peptide is not to provide catalytic groups but to influence catalysis indirectly through stabilization of the ternary Lit⅐Gol⅐EF-Tu complex. Hence, phage-elicited suicide of E. coli K-12 by Lit is a variant form of "cofactor-induced activation," a mechanism of protease activation that has only been documented previously in pathogen subversion of mammalian hemostasis cascades.

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Section: Discussionsupporting

confidence: 71%

Section: Discussionsupporting

confidence: 71%

The Role of an Activating Peptide in Protease-mediated Suicide of Escherichia coli K12

Copeland

Kleanthous

2005

Journal of Biological Chemistry

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“…The polyprotein may even have a different function compared with the mature forms. As an example, several reports have shown that, besides its role in translation elongation, EF-Tu participates in translation arrest (Snyder et al, 2003) and/or ribosome rescue (Wower et al, 2000). In the second case, EF-Tu works together with the S1 protein and SmpB, a transfer-mRNA binding protein, to regulate transfer-mRNA binding to ribosomes.…”

Section: Discussionmentioning

confidence: 99%

Chloroplast Elongation Factor Ts Pro-Protein Is an Evolutionarily Conserved Fusion with the S1 Domain-Containing Plastid-Specific Ribosomal Protein-7

2004

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The components of chloroplast translation are similar to those of prokaryotic translation but contain some additional unique features. Proteomic analysis of the Chlamydomonas reinhardtii chloroplast ribosome identified an S1-like protein, plastid-specific ribosomal protein-7 (PSRP-7), as a stoichiometric component of the 30S subunit. Here, we report that PSRP-7 is part of a polyprotein that contains PSRP-7 on its amino end and two translation elongation factor Ts (EF-Ts) domains at the carboxy end. We named this polyprotein PETs (for polyprotein of EF-Ts). Pets is a single-copy gene containing the only chloroplast PSRP-7 and EF-Ts sequences found in the C. reinhardtii genome. The pets precursor transcript undergoes alternative splicing to generate three mRNAs with open reading frames (ORFs) of 1.68, 1.8, and 3 kb. A 110-kD pro-protein is translated from the 3-kb ORF, and the majority of this protein is likely posttranslationally processed into the 65-kD protein PSRP-7 and a 55-kD EF-Ts. PETs homologs are found in Arabidopsis thaliana and rice (Oryza sativa). The conservation of the 110-kD PETs polyprotein in the plant kingdom suggests that PSRP-7 and EF-Ts function together in some aspects of chloroplast translation and that the PETs pro-protein may have a novel function as a whole

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“…Although Abi systems have been studied predominantly using lactococcal systems, because of their potential economic importance (8) they have been identified in some gram-negative species, such as Escherichia coli, Vibrio cholerae, Shigella dysenteriae, and Erwinia carotovora (9,14,36,38). The prr and lit systems of E. coli have been studied at the molecular level, and their mode of action and mode of activation by incoming phage have been identified (2,37,38). In contrast, lactococcal Abi systems have been characterized mainly by the range of phages actively aborted and the scale of these effects, and the Abi systems have been grouped based on general modes of action (8,12).…”

mentioning

confidence: 99%

Mutagenesis and Functional Characterization of the RNA and Protein Components of the toxIN Abortive Infection and Toxin-Antitoxin Locus of Erwinia

et al. 2009

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Bacteria are constantly challenged by bacteriophage (phage) infection and have developed multiple adaptive resistance mechanisms. These mechanisms include the abortive infection systems, which promote "altruistic suicide" of an infected cell, protecting the clonal population. A cryptic plasmid of Erwinia carotovora subsp. atroseptica, pECA1039, has been shown to encode an abortive infection system. This highly effective system is active across multiple genera of gram-negative bacteria and against a spectrum of phages. Designated ToxIN, this two-component abortive infection system acts as a toxin-antitoxin module. ToxIN is the first member of a new type III class of protein-RNA toxin-antitoxin modules, of which there are multiple homologues crossgenera. We characterized in more detail the abortive infection phenotype of ToxIN using a suite of Erwinia phages and performed mutagenesis of the ToxI and ToxN components. We determined the minimal ToxI RNA sequence in the native operon that is both necessary and sufficient for abortive infection and to counteract the toxicity of ToxN. Furthermore, site-directed mutagenesis of ToxN revealed key conserved amino acids in this defining member of the new group of toxic proteins. The mechanism of phage activation of the ToxIN system was investigated and was shown to have no effect on the levels of the ToxN protein. Finally, evidence of negative autoregulation of the toxIN operon, a common feature of toxin-antitoxin systems, is presented. This work on the components of the ToxIN system suggests that there is very tight toxin regulation prior to suicide activation by incoming phage.

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Regulation of Translation of the Head Protein of T4 Bacteriophage by Specific Binding of EF-Tu to a Leader Sequence

Cited by 10 publications

References 30 publications

The Role of an Activating Peptide in Protease-mediated Suicide of Escherichia coli K12

The Role of an Activating Peptide in Protease-mediated Suicide of Escherichia coli K12

Chloroplast Elongation Factor Ts Pro-Protein Is an Evolutionarily Conserved Fusion with the S1 Domain-Containing Plastid-Specific Ribosomal Protein-7

Mutagenesis and Functional Characterization of the RNA and Protein Components of the toxIN Abortive Infection and Toxin-Antitoxin Locus of Erwinia

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