Biochemical properties of bacterial reverse transcriptase-related (rvt) gene products : multimerization, protein priming, and nucleotide preference
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Cellular reverse transcriptase-related (rvt) genes represent a novel class of reverse transcriptases (RTs), which are only distantly related to RTs of retrotransposons and retroviruses, but, similarly to telomerase RTs, are immobilized in the genome as single-copy genes. They have been preserved by natural selection throughout the evolutionary history of large taxonomic groups, including most fungi, a few plants and invertebrates, and even certain bacteria, being the only RTs present across different domains of life. Bacterial rvt genes are exceptionally rare but phylogenetically related, consistent with common origin of bacterial rvt genes rather than eukaryote-to-bacteria transfer. To investigate biochemical properties of bacterial RVTs, we conducted in vitro studies of recombinant HaRVT protein from the filamentous gliding bacterium Herpetosiphon aurantiacus (Chloroflexi). Although HaRVT does not utilize externally added standard primer-template combinations, in the presence of divalent manganese it can polymerize very short products, using dNTPs rather than NTPs, with a strong preference for dCTP incorporation. Further, we investigated the highly conserved N- and C-terminal domains, which distinguish RVT proteins from other RTs. We show that the N-terminal coiled-coil motif, which is present in nearly all RVTs, is responsible for the ability of HaRVT to multimerize in solution, forming up to octamers. The C-terminal domain may be capable of protein priming, which is abolished by site-directed mutagenesis of the catalytic aspartate and greatly reduced in the absence of the conserved tyrosine residues near the C-terminus. The unusual biochemical properties displayed by RVT in vitro will provide the basis for understanding its biological function in vivo.
Author Posting. © The Author(s), 2018. This is the author's version of the work. It is posted here by permission of Springer for personal use, not for redistribution. The definitive version was published in Current Genetics 64 (2018): 1287-1301, doi:10.1007/s00294-018-0844-6.
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Suggested CitationPreprint: Yushenova, Irina A., Arkhipova, Irina R., "Biochemical properties of bacterial reverse transcriptase-related (rvt) gene products : multimerization, protein priming, and nucleotide preference", 2018-05, https://doi.org/10.1007/s00294-018-0844-6, https://hdl.handle.net/1912/10375
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