High-throughput mutagenesis reveals functional determinants for DNA targeting by activation-induced deaminase.

ABSTRACT: Antibody maturation is a critical immune process governed by the enzyme activation-induced deaminase (AID), a member of the AID/APOBEC DNA deaminase family. AID/APOBEC deaminases preferentially target cytosine within distinct preferred sequence motifs in DNA, with specificity largely conferred by a small 9-11 residue protein loop that differs among family members. Here, we aimed to determine the key functional characteristics of this protein loop in AID and to thereby inform our understanding of the mode of DNA engagement. To this end, we developed a methodology (Sat-Sel-Seq) that couples saturation mutagenesis at each position across the targeting loop, with iterative functional selection and next-generation sequencing. This high-throughput mutational analysis revealed dominant characteri

SUBMITTER: Gajula KS 

PROVIDER: S-EPMC4150791 | biostudies-literature | 2014 Sep

REPOSITORIES: biostudies-literature