TY - JOUR
T1 - An Atypical AAA+ ATPase Assembly Controls Efficient Transposition through DNA Remodeling and Transposase Recruitment
AU - Arias-Palomo, Ernesto
AU - Berger, James M.
N1 - Funding Information:
The authors are grateful to: Eva Nogales and her research group, and the beamline 8.3.1 staff at the Advanced Light Source, for help with structural data collection and analysis; Fang Wu from the Macrolab (UC, Berkeley) for her assistance with the site-directed mutagenesis; Nancy Craig for critically reading the manuscript; and the Berger laboratory for helpful discussions. This work has been supported by a post-doctoral fellowship from the “Programa Nacional de Movilidad de Recursos Humanos del Plan Nacional de I+D+i 2008-2011” from the Spanish Ministry of Education (to E.A.P.), and the NIGMS and Mathers Foundation (GM071747 and 9005-6422 to J.M.B.).
Publisher Copyright:
© 2015 Elsevier Inc.
PY - 2015/8/17
Y1 - 2015/8/17
N2 - Summary Transposons are ubiquitous genetic elements that drive genome rearrangements, evolution, and the spread of infectious disease and drug-resistance. Many transposons, such as Mu, Tn7, and IS21, require regulatory AAA+ ATPases for function. We use X-ray crystallography and cryo-electron microscopy to show that the ATPase subunit of IS21, IstB, assembles into a clamshell-shaped decamer that sandwiches DNA between two helical pentamers of ATP-associated AAA+ domains, sharply bending the duplex into a 180° U-turn. Biochemical studies corroborate key features of the structure and further show that the IS21 transposase, IstA, recognizes the IstB•DNA complex and promotes its disassembly by stimulating ATP hydrolysis. Collectively, these studies reveal a distinct manner of higher-order assembly and client engagement by a AAA+ ATPase and suggest a mechanistic model where IstB binding and subsequent DNA bending primes a selected insertion site for efficient transposition.
AB - Summary Transposons are ubiquitous genetic elements that drive genome rearrangements, evolution, and the spread of infectious disease and drug-resistance. Many transposons, such as Mu, Tn7, and IS21, require regulatory AAA+ ATPases for function. We use X-ray crystallography and cryo-electron microscopy to show that the ATPase subunit of IS21, IstB, assembles into a clamshell-shaped decamer that sandwiches DNA between two helical pentamers of ATP-associated AAA+ domains, sharply bending the duplex into a 180° U-turn. Biochemical studies corroborate key features of the structure and further show that the IS21 transposase, IstA, recognizes the IstB•DNA complex and promotes its disassembly by stimulating ATP hydrolysis. Collectively, these studies reveal a distinct manner of higher-order assembly and client engagement by a AAA+ ATPase and suggest a mechanistic model where IstB binding and subsequent DNA bending primes a selected insertion site for efficient transposition.
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U2 - 10.1016/j.cell.2015.07.037
DO - 10.1016/j.cell.2015.07.037
M3 - Article
C2 - 26276634
AN - SCOPUS:84939215665
SN - 0092-8674
VL - 162
SP - 860
EP - 871
JO - Cell
JF - Cell
IS - 4
M1 - 8352
ER -