Abstract

Rolling-circle replication (RCR) elements constitute a diverse group that includes viruses, plasmids, and transposons, present in hosts from all domains of life. Eukaryotic RCR transposons, also known as Helitrons, are found in species from all eukaryotic kingdoms, sometimes representing a large portion of their genomes. Despite the impact of Helitrons on their hosts, knowledge about their relationship with other RCR elements is still elusive. Here, we compared the endonuclease domain sequence of Helitron transposases with the corresponding region from RCR proteins found in a wide variety of mobile genetic elements. To do that, we used a stepwise alignment approach followed by phylogenetic and multidimensional scaling analyses. Although it has been suggested that Helitrons might have originated from prokaryotic transposons or eukaryotic viruses, our results indicate that Helitron transposases share more similarities with proteins from prokaryotic viruses and plasmids instead. We also provide evidence for the division of RCR endonucleases into three groups (Y1, Y2, and Yx), covering the whole diversity of this protein family. Together, these results point to prokaryotic elements as the likely closest ancestors of eukaryotic RCR transposons, and further demonstrate the fluidity that characterizes the boundaries separating viruses, plasmids, and transposons.

Highlights

  • Rolling-circle replication (RCR) proteins are essential components of many genetic elements found in all three domains of life

  • Their transposition is thought to occur by a mechanism similar to the one proposed for bacterial RCR transposable elements (TEs), like the IS91 family of elements [4,5,6]

  • We investigated the relationship of the Helitron RepHel with other RCR proteins by analyzing the RCRE amino acid sequences from a wide variety of mobile genetic elements, including TEs, plasmids, and viruses

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Summary

Introduction

Rolling-circle replication (RCR) proteins are essential components of many genetic elements found in all three domains of life. The number of catalytic tyrosines has been used to tentatively classify RCR proteins between two superfamilies [17], there is currently no phylogenetic support for this distinction In view of these observations, and considering that domain rearrangements are not uncommon during protein evolution [18], the first scenario (i.e., that Helitrons originated from a prokaryotic ancestral RCR TE) seems to be more parsimonious, as the acquisition of a S1H domain would be the only major evolutionary step in a prokaryotic to eukaryotic RCR TE transition. We investigated the relationship of the Helitron RepHel with other RCR proteins by analyzing the RCRE amino acid sequences from a wide variety of mobile genetic elements, including TEs, plasmids, and viruses. We show that the HUH family of endonucleases can be divided into three major phylogenetic groups comprised of RCR proteins from highly heterogeneous mobile genetic elements

Selecting and Preparing RCRE Domain Sequences
Major RCR Protein Phylogenetic Groups
Helitron Transposase is More Similar to Prokaryotic Proteins
Sequences Retrieval and Selection
Sequence Alignment
NMDS and Phylogenetic Analysis

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