Revision of the nucleotide sequence of the<i>Streptococcus pyogenes</i>plasmid pSM19035<i>rep</i>S gene

Brantl, Sabine; Nowak, A; Behnke, Detlev; Alonso, Juan C.

doi:10.1093/nar/17.23.10110

Cited by 18 publications

(8 citation statements)

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“…Similar differences in the functional importance of individual antisense RNA loops have also been noted for CopA of R1 (33). As (10,11), Sorokin and Khazak (38), and Swinfield et al (39). A, B, a, and b refer to the inverted repeat sequences that are involved in the antisense RNA-driven transcriptional attenuation mechanism.…”

Section: Materlils and Methodssupporting

confidence: 56%

“…A similar situation is true for plasmid pIP501 and its related plasmids pSM19035 and pAMB. The DNA sequences of the replication regions of the latter two plasmids have also been determined (11,38,39), and a comparison revealed a similar organization of transcriptional units (12). All three plasmids carry sequences A/B and a/b as a structural basis for the attenuation mechanism (a compilation of the respective sequences is shown in Fig.…”

Section: Materlils and Methodsmentioning

confidence: 99%

“…Its location coincides with the start point of leading-strand synthesis and the termination point of lagging-strand synthesis that have been mapped to the corresponding sequence on plasmid pAM01 (13). The replication regions of plasmids pIP501, pSM19035, and pAMIB1 have been sequenced (10,11,38,39), and all three were found to code for two is summarized in Fig. 1.…”

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confidence: 93%

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RepR protein expression on plasmid pIP501 is controlled by an antisense RNA-mediated transcription attenuation mechanism

Brantl¹,

Birch-Hirschfeld²,

Behnke³

1993

J Bacteriol

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Expression of the rate-limiting initiator protein RepR of plasmid pIP501 is controlled by the antisense RNAIII. Mutational alteration of individual G residues within the single-stranded loops of RNAIII led to an increase in copy number. In contrast to the G-rich single-stranded loops, two smaller AT-rich loops of RNAIII were found to be dispensable for its inhibitory function. Reciprocal mutations in the same loop compensated for each other's effect, and a destabilization of the major stem structure of RNAIII also resulted in an increased copy number. These data were consistent with the idea that the interaction of RNAIII with its target starts with the formation of a kissing complex between the single-stranded loops of both molecules. The repR mRNA leader sequence, which includes the target of RNAIIIs is able to assume two alternative structures due to the presence of two inverted repeats the individual sequences of which are mutually complementary. In the presence of the antisense RNAIII, one of these inverted repeats (IR2) is forced to fold into a transcriptional terminator structure that prevents transcription of the repR gene. In the absence of RNAIII, formation of the transcriptional terminator is prevented and expression of the essential repR gene can proceed normally. This antisense RNA-driven transcriptional attenuation mechanism was supported by extensive deletional analysis and direct evidence that IR2 functions as a transcriptional terminator.

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Section: Materlils and Methodssupporting

confidence: 56%

Section: Materlils and Methodsmentioning

confidence: 99%

mentioning

confidence: 93%

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RepR protein expression on plasmid pIP501 is controlled by an antisense RNA-mediated transcription attenuation mechanism

Brantl¹,

Birch-Hirschfeld²,

Behnke³

1993

J Bacteriol

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“…No mutation has been demonstrated to explicitly affect the binding process in vitro. Remarkably, under conditions where antisense RNA transcription is com-pletely abolished, no runaway replication is observed [pT181, Carleton et al (1984); pIP501, Brantl and Behnke (1992)]. This is in strong contrast to enterobacterial plasmids, e.g.…”

Section: Discussionmentioning

confidence: 99%

“…pSM19035 (Behnke and Ferretti, 1980). The synthesis of the replication rate-limiting RepR protein is controlled transcriptionally by the repressor CopR (S.Brantl, manuscript submitted) and post-transcriptionally by the antisense RNA, RNAIII (Brantl and Behnke, 1992). Genetic analyses indicated that RNAIII, by interacting with the nascent repR mRNA, RNAII, induces transcriptional termination at an inverted repeat structure located upstream of the repR reading frame (Figure 1;Brantl et al, 1993).…”

Section: Introductionmentioning

confidence: 99%

Antisense RNA-mediated transcriptional attenuation occurs faster than stable antisense/target RNA pairing: an in vitro study of plasmid pIP501.

1994

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Antisense RNA‐mediated transcriptional attenuation is the mode of replication control of several plasmids, among them pIP501. This mechanism implies that the repR mRNAs can fold into two mutually exclusive structures. The formation of one of these structures is induced by binding of the antisense RNA and results in premature termination. Since the fate of the nascent mRNA transcripts depends on the binding rate of the antisense RNA to its target, the control is kinetic. We have studied the antisense RNA, RNAIII, and target RNA, RNAII, whose interaction determines the replication frequency of plasmid pIP501. RNA secondary structures were analyzed using structure‐specific RNases. RNA binding was studied in vitro with normal size and truncated RNAIII species. An in vitro single‐round attenuation assay was developed that permits qualitative and quantitative assessment of inhibition by RNAIII. The effect of varying concentrations of RNAIII species on attenuation was tested and inhibition rate constants were calculated. The inhibition rate constants were at least 10 times higher than the pairing rate constants. Thus, steps preceding stable RNA duplex formation are sufficient to induce RNAIII‐dependent termination of nascent RNAII transcripts.

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Unidirectional theta replication of the structurally stable Enterococcus faecalis plasmid pAM beta 1.

Bruand¹,

Ehrlich²,

Janniere³

1991

The EMBO Journal

112

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Numerous bacterial replicons remain poorly characterized due to difficulties in localization of the replication origin. We have circumvented this problem in the characterization and fine mapping of the origin of plasmid pAM beta 1 by exploiting the Bacillus subtilis termination signal, terC. In terC‐containing derivatives, theta‐form molecules with two invariant endpoints accumulate. The endpoints, which correspond to plasmid origin and terC, were mapped with single‐nucleotide precision. Analysis of the replication intermediates of wild‐type molecules by two‐dimensional gel electrophoresis confirmed the location of the plasmid origin. Our results demonstrate that pAM beta 1 replication proceeds unidirectionally by a theta mechanism. This work confirms the use of termination signals to localize origins, suggests that termination in B. subtilis occurs by a mechanism similar to that of Escherichia coli and establishes that in addition to rolling circle replicating plasmids, Gram positive bacteria harbour plasmids which replicate by a theta mechanism.

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Revision of the nucleotide sequence of theStreptococcus pyogenesplasmid pSM19035repS gene

Cited by 18 publications

References 1 publication

RepR protein expression on plasmid pIP501 is controlled by an antisense RNA-mediated transcription attenuation mechanism

RepR protein expression on plasmid pIP501 is controlled by an antisense RNA-mediated transcription attenuation mechanism

Antisense RNA-mediated transcriptional attenuation occurs faster than stable antisense/target RNA pairing: an in vitro study of plasmid pIP501.

Unidirectional theta replication of the structurally stable Enterococcus faecalis plasmid pAM beta 1.

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