Title |
The H-subunit of the restriction endonuclease CglI contains a prototype DEAD-Z1 helicase-like motor / |
Authors |
Toliušis, Paulius ; Tamulaitienė, Giedrė ; Grigaitis, Rokas ; Tuminauskaitė, Donata ; Šilanskas, Arūnas ; Manakova, Elena ; Venclovas, Česlovas ; Szczelkun, Mark D ; Šikšnys, Virginijus ; Zaremba, Mindaugas |
DOI |
10.1093/nar/gky107 |
Full Text |
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Is Part of |
Nucleic acids research.. Oxford : Oxford University Press. 2018, vol. 46, no 5, p. 2560-2572.. ISSN 0305-1048. eISSN 1362-4962 |
Keywords [eng] |
H-subunit ; Corynebacterium glutamicum ; DEAD-Z1 |
Abstract [eng] |
CglI is a restriction endonuclease from Corynebacterium glutamicum that forms a complex between: two R-subunits that have site specific-recognition and nuclease domains; and two H-subunits, with Su-perfamily 2 helicase-like DEAD domains, and uncharacterized Z1 and C-terminal domains. ATP hydrolysis by the H-subunits catalyses dsDNA translocation that is necessary for long-range movement along DNA that activates nuclease activity. Here, we provide biochemical and molecular modelling evidence that shows that Z1 has a fold distantly-related to RecA, and that the DEAD-Z1 domains together form an ATP binding interface and are the prototype of a previously undescribed monomeric helicase-like motor. The DEAD-Z1 motor has unusual Walker A and Motif VI sequences those nonetheless have their expected functions. Additionally, it contains DEAD-Z1-specific features: an H/H motif and a loop (aa 163-aa 172), that both play a role in the coupling of ATP hydrolysis to DNA cleavage. We also solved the crystal structure of the C-terminal domain which has a unique fold, and demonstrate that the Z1-C domains are the principal DNA binding interface of the H-subunit. Finally, we use small angle X-ray scattering to provide a model for how the H-subunit domains are arranged in a dimeric complex. |
Published |
Oxford : Oxford University Press |
Type |
Journal article |
Language |
English |
Publication date |
2018 |
CC license |
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