Functional coupling of duplex translocation to DNA cleavage in a type I restriction enzyme

Mikalai Lapkouski¹, Tatsiana Baikova^1,2, Ladislav Csefalvay¹, Dhiraj Sinha^1,2, Alina Kevorkova^1,2, Vyas Ramasubramani³, Alena Guzanova⁴, Abdul Samad¹, Morteza Khabiri¹, Pavel Janscak⁵, Ivana Kutá Smatanova¹, Santosh Panjikar⁵, Marie Weiserova⁴, Jannette Carey³,  Eva Csefalvay¹, Rüdiger Ettrich^1,2

¹Institute of Nanobiology and Structural Biology of GCRC, Academy of Sciences of the Czech Republic, Nove Hrady, Czech Republic 

²Faculty Of Sciences, University of South Bohemia, Czech Republic
³Chemistry Department, Princeton University, Princeton, New Jersey, USA
⁴Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic
⁵EMBL Hamburg Outstaion, c/o DESY, Hamburg, Germany 

⁶Institute of Molecular Cancer Research, University of Zurich

The recent structure of the first intact motor subunit of a Type I restriction enzyme suggested a mechanism by which stalled translocation triggers DNA cleavage. Cleavage assays in vivo, ATPase assays in vitro, computational modeling and the crystal structures of active and inactive mutants now reveal how interdomain engagement brings DNA cleavage under the control of translocation.

We gratefully acknowledge support from the Grant Agency of the Czech Republic (P207-12-2323), and joint Czech - US National Science Foundation International Research Cooperation (ME09016 and INT03-09049), Additionally, A.K. was supported by the University of South Bohemia, grant GAJU 170/2010/P.