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Genetic transformation in Streptococcus pneumoniae: nucleotide sequence and predicted amino acid sequence of recP.

ABSTRACT: We present the complete nucleotide sequence of recP, a locus required for high-efficiency recombination of chromosomal DNA during genetic transformation in Streptococcus pneumoniae. The sequence was determined by using plasmid DNA templates and synthetic oligonucleotide primers. The locus contained a single large open reading frame, ORF1, of 1,968 base pairs (bp). ORF1 is included within the recP locus previously mapped genetically and accounts for 94% of its extent. The predicted molecular weight of the largest polypeptide encoded within ORF1, 71,662, coincided with that measured previously (72,000) for the product of in vitro transcription-translation of the cloned recP locus. A Shine-Dalgarno sequence (AGAAAGGA; delta G = -17 kcal [ca. -71.1 kJ]) lay 6 bp upstream of ORF1. A sequence (TTGcat-17 bp-TATAAT) similar to the E. coli sigma-70 promoter consensus was located 52 bp upstream of ORF1. This putative promoter overlapped a structure consisting of two perfect inverted 10-bp repeats and a loop of 8 bp.

SUBMITTER: Radnis BA

PROVIDER: S-EPMC213341 | biostudies-other | 1990 Jul

REPOSITORIES: biostudies-other

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Genetic transformation in Streptococcus pneumoniae: nucleotide sequence and predicted amino acid sequence of recP.

Radnis B A BA Rhee D K DK Morrison D A DA

Journal of bacteriology 19900701 7

We present the complete nucleotide sequence of recP, a locus required for high-efficiency recombination of chromosomal DNA during genetic transformation in Streptococcus pneumoniae. The sequence was determined by using plasmid DNA templates and synthetic oligonucleotide primers. The locus contained a single large open reading frame, ORF1, of 1,968 base pairs (bp). ORF1 is included within the recP locus previously mapped genetically and accounts for 94% of its extent. The predicted molecular weig ...[more]

PMID: 2361942

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Project description:Partial duplication of genetic material is prevalent in eukaryotes and provides potential for evolution of new traits. Prokaryotes, which are generally haploid in nature, can evolve new genes by partial chromosome duplication, known as merodiploidy. Little is known about merodiploid formation during genetic exchange processes, although merodiploids have been serendipitously observed in early studies of bacterial transformation. Natural bacterial transformation involves internalization of exogenous donor DNA and its subsequent integration into the recipient genome by homology. It contributes to the remarkable plasticity of the human pathogen Streptococcus pneumoniae through intra and interspecies genetic exchange. We report that lethal cassette transformation produced merodiploids possessing both intact and cassette-inactivated copies of the essential target gene, bordered by repeats (R) corresponding to incomplete copies of IS861. We show that merodiploidy is transiently stimulated by transformation, and only requires uptake of a ~3-kb DNA fragment partly repeated in the chromosome. We propose and validate a model for merodiploid formation, providing evidence that tandem-duplication (TD) formation involves unequal crossing-over resulting from alternative pairing and interchromatid integration of R. This unequal crossing-over produces a chromosome dimer, resolution of which generates a chromosome with the TD and an abortive chromosome lacking the duplicated region. We document occurrence of TDs ranging from ~100 to ~900 kb in size at various chromosomal locations, including by self-transformation (transformation with recipient chromosomal DNA). We show that self-transformation produces a population containing many different merodiploid cells. Merodiploidy provides opportunities for evolution of new genetic traits via alteration of duplicated genes, unrestricted by functional selective pressure. Transient stimulation of a varied population of merodiploids by transformation, which can be triggered by stresses such as antibiotic treatment in S. pneumoniae, reinforces the plasticity potential of this bacterium and transformable species generally.

Dataset Information

Genetic transformation in Streptococcus pneumoniae: nucleotide sequence and predicted amino acid sequence of recP.

Publications

Genetic transformation in Streptococcus pneumoniae: nucleotide sequence and predicted amino acid sequence of recP.

Similar Datasets

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