Structural Investigations on the Interactions between Cytidine Deaminase Human APOBEC3G and DNA

Human APOBEC3G (A3G) inhibits the replication of human immunodeficiency virus-1 by deaminating cytidine at the 3'-end in the target motif 5'-CCC-3' in viral cDNA during reverse transcription. It in vitro deaminates two consecutive cytidines in a 3'->5' order. Although a crystal structure of the A3G catalytic domain (A3G-CD2) with DNA was reported, it is unknown why residues involved in enzymatic reaction are distributed widely. Here, we introduced an iodine atom into the C-5 position of cytidine (dC₆ ^I ) in DNA 5'-ATTC₄ C₅ C₆ ^I A₇ ATT-3' (TCCC₆ ^I ). It switches the deamination sequence preference from CCC to TCC, although small dC₆ ^I deamination was observed. Solution structures of A3G-CD2 in complexes with products DNA TCUC₆ ^I and TCUU₆ ^I indicate that the substrate DNA binds A3G-CD2 in TCC and CCC modes. The dC₆ deamination correlates with the 4^th base type. The CCC mode favours dC₆ deamination, while the TCC mode results in dC₅ deamination. These studies present an extensive basis to design inhibitors to impede viral evolvability.