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The load of structural evidence favors the idea that an ATP-dependent ligase may be the ancestral state from which NAD+-dependent ligases progressed, by acquisition of the specialized structural area (Ia) that binds the NMN leaving group. All identified NAD+-dependent ligases have area Ia, and they can't use ATP being a substrate for ligase adenylylation, presumably simply because LigA enzymes deficiency “motif IV” on the OB area, the part of ATP-dependent DNA ligases that is believed to interact with the PPi leaving team of ATP (five, 19). This “both/or” circumstance of DNA ligase substrate specificity has been roiled by numerous seemingly conflicting studies concerning the nucleotide specificity of archaeal DNA ligases.

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