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Insight into the recognition mechanism of EcoRV
Author(s) -
Zahran Mai,
Imhof Petra
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.461.25
Subject(s) - ecorv , restriction enzyme , dna , recognition sequence , cleavage (geology) , chemistry , sequence (biology) , enzyme , hindiii , genetics , biochemistry , biology , paleontology , fracture (geology)
EcoRV, a restriction enzyme in Escherichia coli, destroys invading foreign DNA by cleaving it at the center step of a GATATC sequence. In the EcoRV–cognate DNA crystallographic complex, a sharp kink of 50° has been found at the center base‐pair step (TA). In a previous research, we have examined the interplay between the intrinsic propensity of the cognate sequence to kink and the induction by the enzyme, and have understood the mechanism of action of the enzyme and the intrinsically behavior of the cognate sequence to undergo a kink allowing the proper placement of the atoms for cleavage. We have also demonstrated that the enzyme requires 2 magnesium ions at the catalytic site to perform the cleavage. In the present study, we perform multiple independent molecular dynamics simulations to investigate the behavior of a mutant EcoRV (E45A), in the presence of one or two magnesium ions and complexed to three DNA sequences: the cognate sequence, GATATC (TA); the non‐cognate sequence, GAATTC (AT); and with the cognate sequence methylated on the first adenine GA CH3 TATC (TA‐CH 3 ). The results give further insight into the recognition and cleavage mechanism of EcoRV. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .