1887

Abstract

Many Gram-positive and Gram-negative bacteria possess natural competence mechanisms for DNA capture and internalization. In , natural competence is absolutely dependent upon the presence of a seven-gene operon known as the operon (). In species of , this function has been described for a four-gene operon ( in and in ). In this study, a nine- operon (named ) required for natural competence in was identified and characterized. Orf analysis of this operon indicates that the first four Orfs (ComYA–D) share strong homology with ComYA–D of and CglA–D of , the fifth to seventh Orfs (ComYE–G) match conserved hypothetical proteins from various species of with ComYF possessing a predicted ComGF domain, the eighth Orf (ComYH) shows a strong homology to numerous DNA methyltransferases from restriction/modification systems, and the ninth Orf (ComYI) is homologous to acetate kinase (AckA). RT-PCR analysis of the junctions confirmed that all nine s were present in a single transcript, while real-time RT-PCR analysis demonstrated that these s were expressed at a level very similar to that of the first in the operon. Mutations were constructed in all nine putative s. The first seven genes () were found to be essential for natural competence, while and had reduced and normal natural competence ability, respectively. Analyses of –luciferase reporter fusions indicated that expression is growth-phase dependent, with maximal expression at an OD of about 0·2, while mutations in , and reduced the level of expression. In addition, operon expression appears to be correlated with natural competence ability.

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2005-01-01
2024-03-29
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