Structural Basis of hAT Transposon End Recognition by Hermes, an Octameric DNA Transposase from Musca domestica

Hickman, A.B.; Ewis, Hosam E.; Li, Xianghong; Knapp, Joshua A.; Laver, Thomas W; Doss, Anna Louise; Tolun, Gökhan; Steven, Alasdair C.; Grishaev, Alexander; Bax, Ad; Atkinson, Peter W.; Craig, Nancy L.; Dyda, Fred

doi:10.1016/j.cell.2014.05.037

Cited by 67 publications

(146 citation statements)

References 82 publications

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“…RAG2, which works with ZnH2 to embrace coding flank DNAs, is unique in RAG1/2 and absent in other DNA transposases, which undergo a similar two-step DNA double-strand cleavage (Hickman et al, 2014). The role of RAG2 appears to be to equalize DNA binding in the PRC and HFC and support this dramatic DNA transformation.…”

Section: Discussionmentioning

confidence: 99%

Cracking the DNA Code for V(D)J Recombination

Kim

Chuenchor²,

Chen³

et al. 2018

Molecular Cell

117

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Summary To initiate V(D)J recombination for generating the adaptive immune response of vertebrates, RAG1/2 recombinase cleaves DNA at a pair of recombination signal sequences, the 12- and 23-RSS. We have determined crystal and cryoEM structures of RAG1/2 with DNA in the pre-reaction and hairpin-forming complexes up to 2.75Å resolution. Both protein and DNA exhibit structural plasticity and undergo dramatic conformational changes. Coding-flank DNAs rotate, shift and deform extensively for nicking and hairpin formation. Two intertwined RAG1 subunits crisscross four times between the asymmetric pair of severely bent 12/23-RSS DNAs. Location-sensitive bending of 60° and 150° in 12- and 23-RSS spacers, respectively, must occur for RAG1/2 to capture the nonamers and pair the heptamers for symmetric double-strand breakage. DNA pairing is thus sequence-context dependent and structure specific, which partly explains the “beyond 12/23” restriction. Finally, catalysis in crystallo reveals the process of DNA hairpin formation and its stabilization by interleaved base stacking.

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Section: Discussionmentioning

confidence: 99%

Cracking the DNA Code for V(D)J Recombination

Kim

Chuenchor²,

Chen³

et al. 2018

Molecular Cell

117

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“…Mu, Tn7, and Hermes transpososomes are the most complex in requiring multiple transposase subunits (Chaconas & Harshey, 2002, Holder & Craig, 2010, Hickman et al , 2014). In the case of Mu and retroviral integrases, although only two subunits carry out the chemistry of transposition, the stable form of the transpososome is a tetramer (Lavoie et al, 1991, Cherepanov et al , 2011, Montano et al, 2012).…”

Section: Discussionmentioning

confidence: 99%

Controlling DNA degradation from a distance: a new role for the Mu transposition enhancer

Choi

Saha

Jang

et al. 2014

Molecular Microbiology

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Summary Phage Mu is unique among transposable elements in employing a transposition enhancer. The enhancer DNA segment is the site where the transposase MuA binds and makes bridging interactions with the two Mu ends, interwrapping the ends with the enhancer in a complex topology essential for assembling a catalytically active transpososome. The enhancer is also the site at which regulatory proteins control divergent transcription of genes that determine the phage lysis-lysogeny decision. Here we report a third function for the enhancer - that of regulating degradation of extraneous DNA attached to both ends of infecting Mu. This DNA is protected from nucleases by a phage protein until Mu integrates into the host chromosome, after which it is rapidly degraded. We find that leftward transcription at the enhancer, expected to disrupt its topology within the transpososome, blocks degradation of this DNA. Disruption of the enhancer would lead to the loss or dislocation of two non-catalytic MuA subunits positioned in the transpososome by the enhancer. We provide several lines of support for this inference, and conclude that these subunits are important for activating degradation of the flanking DNA. This work also reveals a role for enhancer topology in phage development.

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“…( a ) Hermes (PDB: 4D1Q) 37 , ( b ) bacterial Tn5 (PDB: 1MUS) 40 , and ( c ) RAG1 dimers are shown as ribbon diagrams in two orthogonal views, with the dyad perpendicular to the viewing plane (left) or in the plane (right). Each dimer consists of a cyan and a green subunit.…”

Section: Extended Datamentioning

confidence: 99%

Crystal structure of the V(D)J recombinase RAG1–RAG2

Kim¹,

Lapkouski

Yang³

et al. 2015

Nature

147

196

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Summary V(D)J recombination in the vertebrate immune system generates a highly diverse population of immunoglobulins and T cell receptors by combinatorial joining of segments of coding DNA. The RAG1-RAG2 protein complex (RAG1/2) initiates this site-specific recombination by cutting DNA at specific sites flanking the coding segments. We report here the crystal structure of the RAG1/2 complex at 3.2Å resolution. The 230 kDa RAG1/2 heterotetramer is Y-shaped, with the N-terminal domains of the two RAG1 chains forming an intertwined stalk. Each RAG1/2 heterodimer composes one arm of the “Y” with the active site in the center and RAG2 at its tip. The RAG1/2 structure rationalizes more than 60 mutations identified in immunodeficient patients, as well as a large body of genetic and biochemical data. The architectural similarity between RAG1 and the hairpin-forming transposases Hermes and Tn5 suggests the evolutionary conservation of these DNA rearrangements.

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Structural Basis of hAT Transposon End Recognition by Hermes, an Octameric DNA Transposase from Musca domestica

Cited by 67 publications

References 82 publications

Cracking the DNA Code for V(D)J Recombination

Cracking the DNA Code for V(D)J Recombination

Controlling DNA degradation from a distance: a new role for the Mu transposition enhancer

Crystal structure of the V(D)J recombinase RAG1–RAG2

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