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• W2012631231 abstract "ABSTRACT During V(D)J recombination, RAG targeting to correct sites versus off-target sites relies on both DNA sequence features and on chromatin marks. Kinetic analysis using the first highly active full-length purified RAG1/RAG2 complexes has now allowed us to define the important catalytic features of this complex. We found that the overall rate of nicking, but not hairpinning, is critical for the discrimination between correct (optimal) versus off-target (suboptimal) sites used in human T-cell lymphomas, and we show that the C-terminal portion of RAG2 is required for this. This type of kinetic analysis permits us to analyze only the catalytically active RAG complex, in contrast to all other methods, which are unavoidably confounded by mixture with inactive RAG complexes. Moreover, we can distinguish the two major features of any enzymatic catalysis: the binding constant ( K D ) and the catalytic turnover rate, k cat . Beyond a minimal essential threshold of heptamer quality, further suboptimal heptamer deviations primarily reduce the catalytic rate constant k cat for nicking. Suboptimal nonamers reduce not only the binding of the RAG complex to the recombination site ( K D ) but also the catalytic rate constant, consistent with a tight interaction between the RAG complex and substrate during catalysis. These features explain many aspects of RAG physiology and pathophysiology." @default.
• W2012631231 created "2016-06-24" @default.
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• W2012631231 date "2012-01-15" @default.
• W2012631231 modified "2023-10-14" @default.
• W2012631231 title "Mechanistic Basis for RAG Discrimination between Recombination Sites and the Off-Target Sites of Human Lymphomas" @default.
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• W2012631231 doi "https://doi.org/10.1128/mcb.06187-11" @default.
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