Substrate sequence selectivity of APOBEC3A implicates intra-DNA interactions

5 years ago

Substrate sequence selectivity of APOBEC3A implicates intra-DNA interactions

N., Kurt Yilmaz, Matsuo, B. A., C. A., M., H., S., Silvas, Myint, Kelch, W., T. V., Schiffer, Somasundaran, Hou

The APOBEC3 (A3) family of human cytidine deaminases is renowned for providing a first line of defense against many exogenous and endogenous retroviruses. However, the ability of these proteins to deaminate deoxycytidines in ssDNA makes A3s a double-edged sword. When overexpressed, A3s can mutate endogenous genomic DNA resulting in a variety of cancers. Although the sequence context for mutating DNA varies among A3s, the mechanism for substrate sequence specificity is not well understood. To characterize substrate specificity of A3A, a systematic approach was used to quantify the affinity for substrate as a function of sequence context, length, substrate secondary structure, and pH. We identified the A3A ssDNA binding motif as (T/C)TC(A/G), and found that A3A binds RNA in a sequence specific manner. Furthermore, A3A bound tighter to its substrate binding motif when in a loop compared to linear oligonucleotide. Our results suggest that the A3A affinity and preference for substrate is modulated by the structure of DNA, and not just its chemical identity. Analysis of previously published co-crystal structures of A3A bound to ssDNA in light of the above findings directed the proposal of a new model for the molecular mechanism underlying A3A sequence preference. On a broader scale, the results of this work not only provide key insights into the mechanism of A3's beneficial roles in the cell, especially in viral restriction, but also into A3's deleterious activity such as in the development of cancer.

Publisher URL: http://biorxiv.org/cgi/content/short/176297v1

DOI: 10.1101/176297