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PDBsum entry 4z20
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Hydrolase/DNA
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PDB id
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4z20
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References listed in PDB file
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Key reference
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Title
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Indirect DNA sequence recognition and its impact on nuclease cleavage activity.
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Authors
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A.R.Lambert,
J.P.Hallinan,
B.W.Shen,
J.K.Chik,
J.M.Bolduc,
N.Kulshina,
L.I.Robins,
B.K.Kaiser,
J.Jarjour,
K.Havens,
A.M.Scharenberg,
B.L.Stoddard.
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Ref.
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Structure, 2016,
24,
862-873.
[DOI no: ]
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PubMed id
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Abstract
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LAGLIDADG meganucleases are DNA cleaving enzymes used for genome engineering.
While their cleavage specificity can be altered using several protein
engineering and selection strategies, their overall targetability is limited by
highly specific indirect recognition of the central four base pairs within their
recognition sites. In order to examine the physical basis of indirect sequence
recognition and to expand the number of such nucleases available for genome
engineering, we have determined the target sites, DNA-bound structures, and
central four cleavage fidelities of nine related enzymes. Subsequent
crystallographic analyses of a meganuclease bound to two noncleavable target
sites, each containing a single inactivating base pair substitution at its
center, indicates that a localized slip of the mutated base pair causes a small
change in the DNA backbone conformation that results in a loss of metal
occupancy at one binding site, eliminating cleavage activity.
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