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bioRxiv · 10.1101/235481

Insights into DNA substrate selection by APOBEC3G from structural, biochemical, and functional studies.

Ziegler, S. J.·Liu, C.·Landau, M.·Buzovetsky, O.·Desimmie, B. A.·Zhao, Q.·Sasaki, T.·Burdick, R. C.·Pathak, V. K.·Anderson, K. S.·Xiong, Y.
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Abstract

Human apolipoprotein B mRNA-editing enzyme-catalytic polypeptide-like 3 (A3) proteins are a family of cytidine deaminases that catalyze the conversion of cytidine to uridine in single-stranded DNA (ssDNA). A3 proteins act in the innate immune response to viral infection by mutating the viral ssDNA. One of the most well-studied human A3 family members is A3G, which is a potent inhibitor of HIV-1. Each A3 protein prefers a specific substrate sequence for catalysis - for example, A3G deaminates the third cytidine in the CCCA sequence motif. However, the interaction between A3G and ssDNA is difficult to characterize due to poor solution behavior of the full-length protein and loss of DNA affinity of the truncated protein. Here, we present a novel DNA-anchoring fusion strategy, which we have used to capture an A3G-ssDNA interaction. We characterized an A3G-DNA binding pocket that is important for the enzyme to scan the DNA for its hotspot. The results provide insights into the mechanism by which A3G selects and deaminates its preferred substrates and help define how A3 proteins are tailored to recognize specific DNA sequences. This knowledge contributes to a better understanding of the mechanism of DNA substrate selection by A3G, as well as A3G antiviral activity against HIV-1.

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Ziegler, S. J., Liu, C., Landau, M., Buzovetsky, O., Desimmie, B. A., Zhao, Q., Sasaki, T., Burdick, R. C., Pathak, V. K., Anderson, K. S., Xiong, Y.. 2017-12-16. Insights into DNA substrate selection by APOBEC3G from structural, biochemical, and functional studies.. https://doi.org/10.1101/235481

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