4BEB

MUTANT (K220E) OF THE HSDR SUBUNIT OF THE ECOR124I RESTRICTION ENZYME IN COMPLEX WITH ATP

Replaces:  2Y3T

Summary for 4BEB

• Entry DOI: 10.2210/pdb4beb/pdb
• Related: 4BE7 4BEC
• Descriptor: TYPE I RESTRICTION ENZYME HSDR, MAGNESIUM ION, ADENOSINE-5'-TRIPHOSPHATE (3 entities in total)
• Functional Keywords: hydrolase, dna modification
• Biological source: ESCHERICHIA COLI
• Total number of polymer chains: 4
• Total formula weight: 483241.13

Authors

Csefalvay, E.,Lapkouski, M.,Guzanova, A.,Csefalvay, L.,Baikova, T.,Shevelev, I.,Janscak, P.,Smatanova, I.K.,Panjikar, S.,Carey, J.,Weiserova, M.,Ettrich, R. (deposition date: 2013-03-07, release date: 2014-03-26, Last modification date: 2023-12-20)

Primary citation

Csefalvay, E.,Lapkouski, M.,Guzanova, A.,Csefalvay, L.,Baikova, T.,Shevelev, I.,Bialevich, V.,Shamayeva, K.,Janscak, P.,Kuta Smatanova, I.,Panjikar, S.,Carey, J.,Weiserova, M.,Ettrich, R.
Functional Coupling of Duplex Translocation to DNA Cleavage in a Type I Restriction Enzyme.
Plos One, 10:28700-, 2015

PubMed Abstract: Type I restriction-modification enzymes are multifunctional heteromeric complexes with DNA cleavage and ATP-dependent DNA translocation activities located on motor subunit HsdR. Functional coupling of DNA cleavage and translocation is a hallmark of the Type I restriction systems that is consistent with their proposed role in horizontal gene transfer. DNA cleavage occurs at nonspecific sites distant from the cognate recognition sequence, apparently triggered by stalled translocation. The X-ray crystal structure of the complete HsdR subunit from E. coli plasmid R124 suggested that the triggering mechanism involves interdomain contacts mediated by ATP. In the present work, in vivo and in vitro activity assays and crystal structures of three mutants of EcoR124I HsdR designed to probe this mechanism are reported. The results indicate that interdomain engagement via ATP is indeed responsible for signal transmission between the endonuclease and helicase domains of the motor subunit. A previously identified sequence motif that is shared by the RecB nucleases and some Type I endonucleases is implicated in signaling.

PubMed: 26039067
DOI: 10.1371/JOURNAL.PONE.0128700

Experimental method

X-RAY DIFFRACTION (2.989 Å)