Structures of ISCth4 transpososomes reveal the role&nbsp;of asymmetry in copy-out/paste-in DNA transposition

Kosek, Dalibor; Hickman, Alison B.; Ghirlando, Rodolfo; He, Susu; Dyda, Fred

doi:10.15252/embj.2020105666

Structures of ISCth4 transpososomes reveal the role of asymmetry in copy-out/paste-in DNA transposition

Journal Article · Fri Oct 02 00:00:00 EDT 2020 · EMBO Journal

DOI:https://doi.org/10.15252/embj.2020105666· OSTI ID:1718960

^[1]; Hickman, Alison B. ^[1]; Ghirlando, Rodolfo ^[1]; He, Susu ^[2]; ^[1]

National Institutes of Health (NIH), Bethesda, MD (United States)
National Institutes of Health (NIH), Bethesda, MD (United States); Nanjing Univ. (China)

Copy-out/paste-in transposition is a major bacterial DNA mobility pathway. It contributes significantly to the emergence of antibiotic resistance, often by upregulating expression of downstream genes upon integration. Unlike other transposition pathways, it requires both asymmetric and symmetric strand transfer steps. Here, we report the first structural study of a copy-out/paste-in transposase and demonstrate its ability to catalyze all pathway steps in vitro. X-ray structures of ISCth4 transposase, a member of the IS256 family of insertion sequences, bound to DNA substrates corresponding to three sequential steps in the reaction reveal an unusual asymmetric dimeric transpososome. During transposition, an array of N-terminal domains binds a single transposon end while the catalytic domain moves to accommodate the varying substrates. These conformational changes control the path of DNA flanking the transposon end and the generation of DNA-binding sites. Our results explain the asymmetric outcome of the initial strand transfer and show how DNA binding is modulated by the asymmetric transposase to allow the capture of a second transposon end and to integrate a circular intermediate.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: National Institutes of Health (NIH); USDOE Office of Science (SC), Basic Energy Sciences (BES)

Grant/Contract Number:: W-31109-ENG-38

OSTI ID:: 1718960

Journal Information:: EMBO Journal, Journal Name: EMBO Journal Journal Issue: 1 Vol. 40; ISSN 0261-4189

Publisher:: EMBO PressCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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59 BASIC BIOLOGICAL SCIENCES
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Structures of ISCth4 transpososomes reveal the role of asymmetry in copy-out/paste-in DNA transposition

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