Enterococcus faecalis plasmid pAD1 replication and maintenance.

Abstract

A determinant has been identified which is required for the stable inheritance of a replicon isolated from the pheromone-responsive E. faecalis plasmid pAD1. This determinant, called par, is encoded on a DNA fragment no larger than 457 bp and directs the production of two small RNAs of approximately 250 (RNAI) and 145 (RNAII) nucleotides. Mutations affecting par RNA production also affected plasmid stability, indicating that these RNAs play a central role in par function. Putative transcription start sites were identified within par by DNA sequence analysis, but the sizes of the observed RNAs and the effects of transposon inserts on RNA production are not entirely consistent with predictions based on the hypothetical RNAs produced from these start sites. The absence of clearly functional open reading frames and the presence of various sequence repeat elements within par suggest that these RNAs may not be translated but may perform some structural role in the partition apparatus. The isolated par determinant was capable of producing both RNAs and stabilizing a heterologous vector, indicating that no other pAD1-encoded determinants are required for par function.