Apr 30, 2020

Determinants of Selective Fidelity in Diversity-Generating Retroelements

BioRxiv : the Preprint Server for Biology
S. HandaPartho Ghosh

Abstract

Diversity-generating retroelements (DGRs) diversify proteins to the greatest extent known in the natural world. These elements are found in a wide variety of microbes, including constituents of the microbial dark matter and the human microbiome. Diversification occurs through selective fidelity, in which genetic information in RNA is reverse transcribed to cDNA faithfully for all bases but adenine. We investigated the determinants of selective fidelity in the prototypical Bordetella bacteriophage DGR using an in vitro system that recapitulates this process. We found that the DGR reverse transcriptase bRT and associated protein Avd, along with a specific DGR RNA that contains a template region and flanking functional elements, had a markedly low catalytic efficiency across a template adenine. This was the case even for the correct base pair with an incoming TTP, in agreement with results from other low fidelity polymerases. We identified the C6 substituent of a template purine as a major determinant of selective fidelity, and also identified two bRT amino acids with counterparts in HIV RT based on an in silico model, R74 and I181, whose substitution led to increased fidelity. Our results provide the first elucidation of nucleoba...Continue Reading

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