Base pairing of RNA I with its complementary sequence in the primer precursor inhibits ColE1 replication

Lacatena, Rosa M.; Cesareni, Gianni

doi:10.1038/294623a0

Cited by 210 publications

(104 citation statements)

References 13 publications

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“…The copts mutations could render a region ofthe primer structurally thermosensitive with regard to its ability to interact with RNA I. Recently, the location ofcis-dominant ColEl mutations has supported the idea that the RNA I target site in the primer lies within the region overlapping with RNA I (27,28). Possibly, sequences outside the RNA I overlap region of the primer are also involved in the primer-RNA I interaction.…”

Section: Discussionmentioning

confidence: 99%

Temperature-sensitive copy number mutants of CoIE1 are located in an untranslated region of the plasmid genome.

Wong¹,

Muesing²,

Polisky³

1982

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

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We have isolated two mutant plasmid derivatives of ColEl that exhibit temperature-sensitive replication properties. Both mutants have a normal copy number at 30°C but increase their copy number 30-to 40-fold after a shift in temperature to 42TC. A plasmid-encoded enzyme, &3-actamase (penicillinase, EC 3.5.2.6), undergoes a 30-to 40-fold increase in specific activity concomitant with the increase in plasmid copy number. The cop' phenotype of these mutants is not due to the synthesis of a temperature-sensitive polypeptide. Both mutations are located in an untranslated region of the plasmid genome encoding two overlapping transcripts involved in plasmid replication: a small transcript known as RNA I that acts as a negative control element in replication and a large transcript that has been characterized as the replication primer in vitro. The mutations alter the sequence encoding the primer but lie immediately 5' to the initiating nucleotide of RNA I, in the RNA I promoter region. The possibility that the temperature-dependent plasmid DNA amplification is a consequence of a temperature-sensitive RNA I promoter was tested by inserting the RNA I promoters from the wild-type and mutant plasmids into a plasmid in which galactokinase expression is dependent upon an exogenous promoter. These experiments demonstrate that the mutant promoters are not temperature-sensitive. Rather, the mutations may affect the secondary structure of the replication primer in a region important for RNA I interaction.Extrachromosomal elements such as the plasmid ColEl are model systems for the study of DNA replication control mechanisms because of their small size, genetic dispensibility, and the fact that they encode information which plays a role in the maintenance of a characteristic and stable copy number (1). Mutations in ColEl derivatives that display elevated copy numbers (designated cop-) have been isolated (2, 3). These mutants increase the copy number from about 15 copies per chromosome to 200-300 per chromosome (2, 3). The behavior of the recessive copj plasmid pOP1A6 was consistent with the existence ofa diffusible, replicon-specific inhibitor ofplasmid DNA replication. We have shown that the mutation in pOP1A6 is a single base pair (bp) change in a region encoding a small, nontranslated RNA molecule designated RNA I (4). RNA I is 108 nucleotides long and is transcribed from a region 450 bp from the origin of replication (5).The simple view that RNA I was a trans-acting, replicon-specific repressor was complicated by the discovery that the region of the genome encoding RNA I also encodes a RNA primer of DNA replication in vitro (6). Thus, the mutation described above alters the sequence of both RNA I and the in vitro replication primer. This primer is 555 nucleotides long and is transcribed from the DNA strand complementary to that encoding RNA I. Synthesis ofthe primer transcript initiates near the termination site of RNA I. Using an in vitro DNA replication system, Itoh and Tomizawa (6) have shown that generation of pri...

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Section: Discussionmentioning

confidence: 99%

Temperature-sensitive copy number mutants of CoIE1 are located in an untranslated region of the plasmid genome.

Wong¹,

Muesing²,

Polisky³

1982

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

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“…For the ColEl plasmid it has been postulated that the tRNA-like cloverleaf structure of RNA I aids in the binding ofRNA I to its target, RNA II (22 …”

Section: Resultsmentioning

confidence: 99%

Protection against tobacco mosaic virus in transgenic plants that express tobacco mosaic virus antisense RNA.

Powell

Stark

Sanders

et al. 1989

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

111

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Transgenic tobacco plants that express RNA sequences complementary to the tobacco mosaic virus (TMV) coat protein (CP) coding sequence with or without the tRNAlike structure at the 3' end of the TMV RNA were produced. Progeny of self-pollinated plants were challenged with TMV to determine their resistance to infection. Plants that expressed RNA sequences complementary to the CP coding region and the 3' untranslated region, including the tRNA-like sequences, were protected from infection by TMV at low levels of inocu- (6) and the expression of the insertion element ISJO (7). In these examples the antisense RNA presumably anneals with its corresponding mRNA and prevents translation (7,8).It has been shown that replication ofthe RNA coliphage SP can be inhibited by antisense RNA by expressing various antisense viral sequences in the host (9, 10). The antisense RNA for the maturase gene was the most effective molecule in these replication inhibition assays, and antisense RNA complementary to sequences at the 3' end of the viral RNA had a lesser effect. Since the latter region contained only the 3' one-third ofthe replicase gene and the 3' noncoding region, the result indicates that antisense RNA may be effective at blocking replication in RNA viruses independent of its ability to block translation. (Fig. 1B)

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“…Naturally occurring asRNAs were first observed in bacteria more than 30 years ago (38,47) and were postulated even earlier for bacteriophage (85). In archaea, the first case of the antisense control of gene expression was reported in 1993 for the extremely halophilic Halobacterium salinarium, lysogenic for phage H, with an asRNA complementary to the first 151 nucleotides (nt) of the transcript T1 (88).…”

Section: Introductionmentioning

confidence: 99%

cis-Antisense RNA, Another Level of Gene Regulation in Bacteria

Georg

Hess

2011

Microbiol Mol Biol Rev

361

348

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SUMMARY A substantial amount of antisense transcription is a hallmark of gene expression in eukaryotes. However, antisense transcription was first demonstrated in bacteria almost 50 years ago. The transcriptomes of bacteria as different as Helicobacter pylori , Bacillus subtilis , Escherichia coli , Synechocystis sp. strain PCC6803, Mycoplasma pneumoniae , Sinorhizobium meliloti , Geobacter sulfurreducens , Vibrio cholerae , Chlamydia trachomatis , Pseudomonas syringae , and Staphylococcus aureus have now been reported to contain antisense RNA (asRNA) transcripts for a high percentage of genes. Bacterial asRNAs share functional similarities with trans -acting regulatory RNAs, but in addition, they use their own distinct mechanisms. Among their confirmed functional roles are transcription termination, codegradation, control of translation, transcriptional interference, and enhanced stability of their respective target transcripts. Here, we review recent publications indicating that asRNAs occur as frequently in simple unicellular bacteria as they do in higher organisms, and we provide a comprehensive overview of the experimentally confirmed characteristics of asRNA actions and intimately linked quantitative aspects. Emerging functional data suggest that asRNAs in bacteria mediate a plethora of effects and are involved in far more processes than were previously anticipated. Thus, the functional impact of asRNAs should be considered when developing new strategies against pathogenic bacteria and when optimizing bacterial strains for biotechnology.

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Base pairing of RNA I with its complementary sequence in the primer precursor inhibits ColE1 replication

Cited by 210 publications

References 13 publications

Temperature-sensitive copy number mutants of CoIE1 are located in an untranslated region of the plasmid genome.

Temperature-sensitive copy number mutants of CoIE1 are located in an untranslated region of the plasmid genome.

Protection against tobacco mosaic virus in transgenic plants that express tobacco mosaic virus antisense RNA.

cis-Antisense RNA, Another Level of Gene Regulation in Bacteria

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