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We identified a functional single strand origin of replication ( sso ) in the integrative and conjugative element ICE Bs1  of  Bacillus subtilis . Integrative and conjugative elements (ICEs, also known as conjugative transposons) are DNA elements typically found integrated into a bacterial chromosome where they are transmitted to daughter cells by chromosomal replication and cell division. Under certain conditions, ICEs become activated and excise from the host chromosome and can transfer to neighboring cells via the element-encoded conjugation machinery. Activated ICE Bs1  undergoes autonomous rolling circle replication that is needed for the maintenance of the excised element in growing and dividing cells. Rolling circle replication, used by many plasmids and phages, generates single-stranded DNA (ssDNA). In many cases, the presence of an  sso  enhances the conversion of the ssDNA to double-stranded DNA (dsDNA) by enabling priming of synthesis of the second DNA strand. We initially identified  sso1  in ICE Bs1  based on sequence similarity to the  sso  of an RCR plasmid. Several functional assays confirmed Sso activity. Genetic analyses indicated that ICE Bs1  uses  sso1  and at least one other region for second strand DNA synthesis. We found that Sso activity was important for two key aspects of the ICE Bs1  lifecycle: 1) maintenance of the plasmid form of ICE Bs1  in cells after excision from the chromosome, and 2) stable acquisition of ICE Bs1  following transfer to a new host. We identified sequences similar to known plasmid  sso 's in several other ICEs. Together, our results indicate that many other ICEs contain at least one single strand origin of replication, that these ICEs likely undergo autonomous replication, and that replication contributes to the stability and spread of these elements.

Identification of a Single Strand Origin of Replication in the Integrative and Conjugative Element ICEBs1 of Bacillus subtilis