SGI1 excludes IncA and IncC plasmids

Abstract

SGI1 and its many variant forms are integrative mobilizable elements that rely on IncA or IncC plasmids for transfer functions. However, the coexistence of SGI1 with the plasmid is unstable in the longer term. Here, we have investigated the effect of SGI1 type integrative elements on the initial entry of these plasmids. Using two transfer proficient IncC plasmids and the IncA plasmid RA1, exclusion indices were 40–100-fold for SGI1-I or SGI1-D which have a complete backbone. Using the SGI1-K and SGI1-LK1 variants that lack backbone segments, loss of a region of 5793 bp that includes the traHG transfer genes and the downstream open reading frame S010 was found to abolish exclusion. S010 was shown to be co-transcribed with traHG and hence also under the control of an AcaDC inducible promoter. However, complementation with a 5.2 kbp fragment that included the traHG-S010 operon did not restore exclusion activity to SGI1-LK1. Part of S013 that encodes a small polypeptide of unknown function, was also lost from SGI1-LK1. S013 and the adjacent S014 gene were also co-transcribed. However complementation with S013-S014 did not restore exclusion activity to SGI1-LK1. Hence, the precise cause of the SGI1-mediated plasmid exclusion remains elusive.