The N-terminal domain of MuB protein has striking structural similarity to DNA-binding domains and mediates MuB filament–filament interactions

Dramićanin, Marija; López-Méndez, Blanca; Boskovic, Jasminka; Campos-Olivas, Ramón; Ramón‐Maiques, Santiago

doi:10.1016/j.jsb.2015.07.004

Cited by 5 publications

(4 citation statements)

References 49 publications

(54 reference statements)

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“…Our results suggest how transposition targeting mechanisms might activate transposition: their ability to present pre-deformed or pre-bent DNA. The structural data available for MuB and IstB (the related targeting ATPase from IS21elements) suggest that they have the ability change the helical conformation of the DNA they coat and/or to form bundles of protein-DNA filaments (Arias-Palomo and Berger, 2015; Dramićanin et al, 2015; Mizuno et al, 2013). Loops could be extruded from these large protein-DNA complexes by transient dissociation events within a filament or as the DNA crosses from one collinear filament to an adjacent one within a bundle.…”

Section: Discussionmentioning

confidence: 99%

Target DNA bending by the Mu transpososome promotes careful transposition and prevents its reversal

Fuller

Rice

2017

eLife

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The transposition of bacteriophage Mu serves as a model system for understanding DDE transposases and integrases. All available structures of these enzymes at the end of the transposition reaction, including Mu, exhibit significant bends in the transposition target site DNA. Here we use Mu to investigate the ramifications of target DNA bending on the transposition reaction. Enhancing the flexibility of the target DNA or prebending it increases its affinity for transpososomes by over an order of magnitude and increases the overall reaction rate. This and FRET confirm that flexibility is interrogated early during the interaction between the transposase and a potential target site, which may be how other DNA binding proteins can steer selection of advantageous target sites. We also find that the conformation of the target DNA after strand transfer is involved in preventing accidental catalysis of the reverse reaction, as conditions that destabilize this conformation also trigger reversal.DOI: http://dx.doi.org/10.7554/eLife.21777.001

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Section: Discussionmentioning

confidence: 99%

Target DNA bending by the Mu transpososome promotes careful transposition and prevents its reversal

Fuller

Rice

2017

eLife

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“…The B protein of Mu (MuB), a non-specific DNA-binding protein and AAA+ ATPase, is essential for the efficient capture and delivery of the target to the transpososome via MuB-MuA interaction; MuB also plays critical roles at all stages of transposition by allosterically activating MuA (see [ 3 , 9 ]). MuB forms ATP-dependent helical filaments, with or without DNA [ 10 – 12 ]. Because of a mismatch between the helical parameters of the MuB filament and that of the bound DNA, it has been proposed that the DNA at the boundary of the MuB filament deforms, creating a DNA bend favored by MuA as a target [ 11 , 13 , 14 ].…”

Section: Introductionmentioning

confidence: 99%

Deep sequencing reveals new roles for MuB in transposition immunity and target-capture, and redefines the insular Ter region of E. coli

Walker

Harshey

2020

Mobile DNA

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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.

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“…The B protein of Mu (MuB), a non-speci c DNA-binding protein and AAA+ ATPase, is essential for the e cient capture and delivery of the target to the transpososome via MuB-MuA interaction; MuB also plays critical roles at all stages of transposition by allosterically activating MuA (see [3,9]). MuB forms ATP-dependent helical laments, with or without DNA [10][11][12]. Because of a mismatch between the helical parameters of the MuB lament and that of the bound DNA, it has been proposed that the DNA at the boundary of the MuB lament deforms, creating a DNA bend favored by MuA as a target [11,13,14].…”

Section: Introductionmentioning

confidence: 99%

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

Walker

Harshey

2020

Preprint

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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.

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The N-terminal domain of MuB protein has striking structural similarity to DNA-binding domains and mediates MuB filament–filament interactions

Cited by 5 publications

References 49 publications

Target DNA bending by the Mu transpososome promotes careful transposition and prevents its reversal

Target DNA bending by the Mu transpososome promotes careful transposition and prevents its reversal

Deep sequencing reveals new roles for MuB in transposition immunity and target-capture, and redefines the insular Ter region of E. coli

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

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