The fourth type of replicon are the repABC-type operons, which ar

The fourth type of replicon are the repABC-type operons, which are specifically found in Alphaproteobacteria and which can be differentiated from the three other groups as in this system the oriV is situated within the replicase gene (Petersen, Selleck Dabrafenib 2011). The comparison of the organization of the three relevant genes on the plasmids from sphingomonads demonstrated that the three groups of ‘megaplasmids’ identified in the course of the sequence comparisons of the individual rep and par genes also corresponded with the gene organization. Thus, in the group of plasmids consisting of pNL1,

pCAR3, pSWIT02 and Mpl (‘Mega-RepAC’), the repA genes are always transcribed in the opposite direction to the parAB genes (Fig. 3). For pNL1 and pCAR3, it has been previously shown that in the sequence space between the repA and parA genes, several 16–17 bp long repeats are present. This indicates that these repeats function as iterons

and thus are the DNA sequences to which the RepA proteins bind (Romine et al., 1999; Shintani et al., 2007). A search for similar iterons at the corresponding position of plasmid pSWIT02 (using the program repfind; Kinase Inhibitor Library http://zlab.bu.edu/repfind) identified three copies of a 14 bp DNA sequence, which was part of the 16 bp iteron found at the respective site in pCAR3. Thus, it can be concluded that the plasmids belonging to the ‘Mega-RepAC-family’ belong to the RepA-group of Alphaproteobacterial replicons as previously defined by Petersen (2011). The ‘Mega-RPA’ group (consisting of plasmids pNL2, pISP1 and Lpl) demonstrated the gene order parA, parB, repA (Fig. 3).

This MYO10 is the same gene order as found in the repABC operons. Unfortunately, the nomenclature of the genes participating in the repABC operons is different from the nomenclature used for the three other types of replicons. Thus, in the case of the repABC operons, RepA and RepB have sequence similarities to proteins involved in active segregation of plasmids – and thus are equivalent to ParA and ParB in the other systems – and RepC is the replication initiator protein – and thus is equivalent to RepA in the other systems (Cevallos et al., 2008). The repABC plasmids show in addition to the highly conserved gene order also further characteristics. Thus, it had been shown that in the large intergenic sequence between repB and repC, a gene is present which codes for a small antisense RNA which is involved in the control of plasmid replication (Cevallos et al., 2008). Therefore, the sequence space between the genes coding for the parB and repA genes of plasmids pNL2, pISP1 and Lpl were analysed and compared with the respective sequences encoding for the antisense RNAs from various plasmids belonging to the repABC family (Venkova-Canoca et al., 2004), but no significant sequence homologies were detected. This suggested that the plasmids of ‘Mega-RPA-group’ do not belong to the repABC plasmids.

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