Deep Sequencing Reveals New Roles for MuB in Transposition Immunity and Target-capture, and Redefines the Insular Ter Region of E. coli

Abstract

Background The target capture protein MuB is responsible for the high efficiency of phage Mu transposition within the E. coli genome. However, some targets are off-limits, such as regions immediately outside the Mu ends (cis-immunity) as well as the entire ~37 kb genome of Mu (Mu genome immunity). Paradoxically, MuB is responsible for cis-immunity and is also implicated in Mu genome immunity, but via different mechanisms. This study was undertaken to dissect the role of MuB in target choice in vivo.Results We tracked Mu transposition from six different starting locations on the E. coli genome, in the presence and absence of MuB. The data reveal that Mu’s ability to sample the entire genome during a single hop in a clonal population is independent of MuB, and that MuB is responsible for cis-immunity, plays a minor role in Mu genome immunity, and facilitates insertions into transcriptionally active regions. Unexpectedly, transposition patterns in the absence of MuB have helped extend the boundaries of the insular Ter segment of the E. coli genome.Conclusions The results in this study demonstrate unambiguously the operation of two distinct mechanisms of Mu target immunity, only one of which is wholly dependent on MuB. The study also reveals several interesting and hitherto unknown aspects of Mu target choice in vivo, particularly the role of MuB in facilitating the capture of promoter and translation start site targets, likely by displacing macromolecular complexes engaged in gene expression. So also, MuB facilitates transposition into the restricted Ter region of the genome.

The bacteriophage mu lysis system–A new mechanism of host lysis?

“…Interestingly, Mu is the most efficient transposable element known (Mizuuchi, 1983;Walker and Harshey, 2020) and can infect and lyse several hosts (Escherichia coli, Shigella sonnei, Citrobacter freundii, Erwinia, and Enterobacter) (Paolozzi and Ghelardini, 2006). Moreover, Mu-like phages were discovered in a wide range of Gramnegative and Gram-positive bacteria (Hull et al, 1978;Hayashi et al, 2001;Toussaint, 2013).…”