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Prediction and functional analysis of the replication origin of the linear plasmid pSCL2 in Streptomyces clavuligerus

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Abstract:

pSCL2 (120 kb), one of the linear plasmids found in Streptomyces clavuligerus NRRL3585, was isolated and partially sequenced. Computational analysis of the central region of pSCL2 revealed the presence of two open reading frames that appear to encode proteins highly homologous to RepL1 and RepL2, replication proteins from pSLA2-L, the large linear plasmid in Streptomyces rochei. The S. clavuligerus open reading frames were designated repC1 and repC2, encoding the proteins RepC1 (150 amino acids) and RepC2 (102 amino acids), respectively. The RepC and RepL proteins have identical translation features and very similar predicted secondary and tertiary structures. Functional analysis confirmed that RepC1 is essential for replication initiation of pSCL2, whereas RepC2 is dispensable but may play a role in copy number control. The RepC and RepL proteins do not show similarity to any other bacterial plasmid replication proteins. Three regions of DNA sequence, Box 1 (1050–850 bp), Box 2 (723–606 bp), and Box 3 (224–168 bp), located upstream of repC1, were also shown to be essential or very important for replication of pSCL2.Key words: pSCL2, Streptomyces clavuligerus, replication origin.

pSCL2 (120 kb), l'un des plasmides linéaires retrouvés chez Streptomyces clavuligerus NRRL3585, fut isolé et partiellement séquencé. Une analyse informatique de la région centrale de pSCL2 a révélé la présence de deux cadres de lecture ouverts semblant coder des protéines hautement homologues à RepL1 et RepL2, des protéines de réplication de pSLA2-L, le grand plasmide linéaire de Streptomyces rochei. Les cadres de lecture ouverts de S. clavuligerus furent désignés repC1 et repC2, codant respectivement les protéines RepC1 (150 acides aminés) et RepC2 (102 acides aminés). Les protéines RepC et RepL avaient des attributs traductionnels identiques et des structures secondaires et tertiaires prédites très semblables. Une analyse fonctionnelle a confirmé que RepC1 est essentiel à l'initiation de la réplication de pSCL2, alors que RepC1 est dispensable mais pourrait jouer un rôle dans le contrôle du nombre de copies. Les protéines RepC et RepL ne démontrent pas de similitude avec aucune autre protéine bactérienne de réplication plasmidique. Trois régions de séquence d'ADN, la Boîte 1 (1050–850 pb), la Boîte 2 (723–606 pb) et la Boîte 3 (224–168 pb), situées en amont de repC1 ont également démontré une contribution essentielle ou très importante à la réplication de pSCL2.Mots clés : pSCL2, Streptomyces clavuligerus, origine de réplication.[Traduit par la Rédaction]

Document Type: Research Article

Publication date: 2006-04-01

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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