Frameshifting by transcriptional slippage is involved in production of MxiE, the transcription activator regulated by the activity of the type III secretion apparatus in <i>Shigella flexneri</i>

Penno, Christophe; Sansonetti, Philippe J.; Parsot, Claude

doi:10.1111/j.1365-2958.2004.04530.x

Cited by 41 publications

(43 citation statements)

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“…CTS1 contains all of the components that have been determined to be necessary for a functional T6SS in other bacteria (95). Interestingly, the icmF homologue in CTS1 has a frameshift at a region containing a polyadenosine tract, which may allow regulation of the gene via transcriptional slippage in an manner analogous to that of mixE of Shigella or the pyrBI locus in E. coli (52,74).…”

Section: T6ssmentioning

confidence: 99%

The Citrobacter rodentium Genome Sequence Reveals Convergent Evolution with Human Pathogenic Escherichia coli

et al. 2010

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Citrobacter rodentium (formally Citrobacter freundii biotype 4280) is a highly infectious pathogen that causes colitis and transmissible colonic hyperplasia in mice. In common with enteropathogenic and enterohemorrhagic Escherichia coli (EPEC and EHEC, respectively), C. rodentium exploits a type III secretion system (T3SS) to induce attaching and effacing (A/E) lesions that are essential for virulence. Here, we report the fully annotated genome sequence of the 5.3-Mb chromosome and four plasmids harbored by C. rodentium strain ICC168. The genome sequence revealed key information about the phylogeny of C. rodentium and identified 1,585 C. rodentium-specific (without orthologues in EPEC or EHEC) coding sequences, 10 prophage-like regions, and 17 genomic islands, including the locus for enterocyte effacement (LEE) region, which encodes a T3SS and effector proteins. Among the 29 T3SS effectors found in C. rodentium are all 22 of the core effectors of EPEC strain E2348/69. In addition, we identified a novel C. rodentium effector, named EspS. C. rodentium harbors two type VI secretion systems (T6SS) (CTS1 and CTS2), while EHEC contains only one T6SS (EHS). Our analysis suggests that C. rodentium and EPEC/EHEC have converged on a common host infection strategy through access to a common pool of mobile DNA and that C. rodentium has lost gene functions associated with a previous pathogenic niche.

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Section: T6ssmentioning

confidence: 99%

The Citrobacter rodentium Genome Sequence Reveals Convergent Evolution with Human Pathogenic Escherichia coli

et al. 2010

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“…The two genes are organized in a single highly conserved bicistronic operon. Although phoN2 encodes a periplasmic protein (5), their expression is regulated by the VirF-VirB cascade (35) and by MxiE in concert with IpgC so that they are expressed in a VirB-dependent, MxiE-independent manner under conditions of nonactivated secretion and are up-regulated, in a MxiEdependent manner, under conditions of activated secretion (8,17,20,26,30), indicating that these genes may be important in postinvasion events related to virulence.phoN2 encodes apyrase, a periplasmic enzyme which belongs to the family of ATP-hydrolyzing enzymes (3, 5, 18) able to sequentially hydrolyze nucleoside triphosphates to diphosphates and then to monophosphates. However, it is not active against monophosphates, a distinguishing feature from acid phosphatases (1,3,41).…”

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Apyrase, the Product of the Virulence Plasmid-Encoded phoN2 ( apy ) Gene of Shigella flexneri , Is Necessary for Proper Unipolar IcsA Localization and for Efficient Intercellular Spread

Santapaola

Chierico²,

Petrucca

et al. 2006

J Bacteriol

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The role in virulence of the Shigella flexneri ospB-phoN2 operon has been evaluated. Here we confirm that OspB is an effector and show that apyrase, the product of phoN2, may be a virulence factor, since it is required for efficient intercellular spreading. Apyrase may be important in a deoxynucleoside triphosphate-hydrolyzing activity-independent manner, suggesting that it may act as an interaction partner in the process of IcsA localization.ospB and phoN2 (apy) are genes located on the virulence plasmid (pINV) of Shigella species and of related enteroinvasive Escherichia coli (EIEC) strains (3,6,35). Indirect evidence indicates that ospB and phoN2 may contribute to pathogenicity (1,3,5,8,17,23,25,35). ospB encodes a protein of unknown function (OspB) surmised to be an effector secreted by the type III secretion (TTS) apparatus of Shigella flexneri (6,20). The two genes are organized in a single highly conserved bicistronic operon. Although phoN2 encodes a periplasmic protein (5), their expression is regulated by the VirF-VirB cascade (35) and by MxiE in concert with IpgC so that they are expressed in a VirB-dependent, MxiE-independent manner under conditions of nonactivated secretion and are up-regulated, in a MxiEdependent manner, under conditions of activated secretion (8,17,20,26,30), indicating that these genes may be important in postinvasion events related to virulence.phoN2 encodes apyrase, a periplasmic enzyme which belongs to the family of ATP-hydrolyzing enzymes (3, 5, 18) able to sequentially hydrolyze nucleoside triphosphates to diphosphates and then to monophosphates. However, it is not active against monophosphates, a distinguishing feature from acid phosphatases (1,3,41). Due to its catalytic activity and primary structure (apyrase presents an exposed N-terminal poly-proline sequence), it has been implicated in the decrease of host cell intracellular ATP, a characteristic metabolic event following S. flexneri infection, as well as in the S. flexneri-induced actin-polymerization process (1,3,5,25,35). Thus, these findings strongly suggest that OspB and apyrase may be virulence factors. The purpose of this study was to investigate the role of both genes in the mechanism of pathogenicity of S. flexneri. OspB is an effector secreted by the TTS apparatus of S. flexneri. To confirm and extend previous reports indicatingOspB as an effector secreted by the TTS apparatus (6, 20), we transduced a nonpolar deletion encompassing mxiA into the S. flexneri M90T derivative strain HND549 (ospB::3ϫFLAG), thus generating HND5311 (mxiA ospB::3ϫFLAG) ( Table 1). C-terminal 3ϫ FLAG tagging was achieved essentially as described by Uzzau et al. (42) using the specific primer pair shown in Table S1 in the supplemental material. Whole-cell extracts and supernatants of exponentially growing bacteria were analyzed by immunoblotting using monoclonal antibodies against the 3ϫ FLAG epitope (Sigma) and mouse polyclonal antibodies preparation directed against apyrase (Fig. 1). Apyrase antiserum was obtained by immunizing BALB/...

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“…In other bacteria, counterpart slippage is required for synthesis of the fulllength rather than the truncated product (2). Disease significant homopolymeric transcriptional slippage occurs in expression of the secretion system of Shigella flexneri (3,4), Citrobacter, and Yersinia (5). Mechanistically similar slippage is involved in several bacterial IS element transposases (6) and probably in Staphyloccocus aureus mapW (7).…”

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Productive mRNA stem loop-mediated transcriptional slippage: Crucial features in common with intrinsic terminators

Penno

Sharma

Coakley

et al. 2015

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

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Escherichia coli and yeast DNA-dependent RNA polymerases are shown to mediate efficient nascent transcript stem loop formation-dependent RNA-DNA hybrid realignment. The realignment was discovered on the heteropolymeric sequence T5C5 and yields transcripts lacking a C residue within a corresponding U5C4. The sequence studied is derived from a Roseiflexus insertion sequence (IS) element where the resulting transcriptional slippage is required for transposase synthesis. The stability of the RNA structure, the proximity of the stem loop to the slippage site, the length and composition of the slippage site motif, and the identity of its 3′ adjacent nucleotides (nt) are crucial for transcripts lacking a single C. In many respects, the RNA structure requirements for this slippage resemble those for hairpin-dependent transcription termination. In a purified in vitro system, the slippage efficiency ranges from 5% to 75% depending on the concentration ratios of the nucleotides specified by the slippage sequence and the 3′ nt context. The only previous proposal of stem loop mediated slippage, which was in Ebola virus expression, was based on incorrect data interpretation. We propose a mechanical slippage model involving the RNAP translocation state as the main motor in slippage directionality and efficiency. It is distinct from previously described models, including the one proposed for paramyxovirus, where following random movement efficiency is mainly dependent on the stability of the new realigned hybrid. In broadening the scope for utilization of transcription slippage for gene expression, the stimulatory structure provides parallels with programmed ribosomal frameshifting at the translation level.transcriptional realignment | stem loop stimulator | heteropolymeric slippage-prone motifs | frameshifting A relatively neglected aspect of gene expression is functionally important RNA-DNA hybrid realignment within the RNA polymerase (RNAP) transcribing coding sequence. Such realigned polymerases yield transcripts lacking, or containing, one or more additional base(s) corresponding to the slippage-prone sequence. The fraction of the mRNA population containing deletions or insertions with respect to the template that are not of 3 nucleotides (nt) or multiples thereof, yields proteins that are either truncated, have C-terminal regions not encoded by the zero frame of the template DNA, or both. Despite the limited investigations, several diverse functional slippage-derived products have already been characterized.Bacterial multisubunit DNA-dependent RNAPs undergo slippage on homopolymeric runs of As or Ts. Transcriptional slippage on a run of nine As in the Thermus thermophilus dnaX gene results in heterogeneous transcripts that lead to 50% of the ribosomes quickly terminating. This termination yields a DNA polymerase subunit that lacks C-terminal domains and is synthesized in a 1:1 ratio with the full-length product (1). In other bacteria, counterpart slippage is required for synthesis of the fulllength rather than the truncat...

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Frameshifting by transcriptional slippage is involved in production of MxiE, the transcription activator regulated by the activity of the type III secretion apparatus in Shigella flexneri

Cited by 41 publications

References 40 publications

The Citrobacter rodentium Genome Sequence Reveals Convergent Evolution with Human Pathogenic Escherichia coli

The Citrobacter rodentium Genome Sequence Reveals Convergent Evolution with Human Pathogenic Escherichia coli

Apyrase, the Product of the Virulence Plasmid-Encoded phoN2 ( apy ) Gene of Shigella flexneri , Is Necessary for Proper Unipolar IcsA Localization and for Efficient Intercellular Spread

Productive mRNA stem loop-mediated transcriptional slippage: Crucial features in common with intrinsic terminators

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