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      Genetic Diversity of Potassium Ion Channel Proteins Encoded by Chloroviruses That Infect Chlorella heliozoae

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          Abstract

          Chloroviruses are large, plaque-forming, dsDNA viruses that infect chlorella-like green algae that live in a symbiotic relationship with protists. Chloroviruses have genomes from 290 to 370 kb, and they encode as many as 400 proteins. One interesting feature of chloroviruses is that they encode a potassium ion (K +) channel protein named Kcv. The Kcv protein encoded by SAG chlorovirus ATCV-1 is one of the smallest known functional K + channel proteins consisting of 82 amino acids. The Kcv ATCV-1 protein has similarities to the family of two transmembrane domain K + channel proteins; it consists of two transmembrane α-helixes with a pore region in the middle, making it an ideal model for studying K + channels. To assess their genetic diversity, kcv genes were sequenced from 103 geographically distinct SAG chlorovirus isolates. Of the 103 kcv genes, there were 42 unique DNA sequences that translated into 26 new Kcv channels. The new predicted Kcv proteins differed from Kcv ATCV-1 by 1 to 55 amino acids. The most conserved region of the Kcv protein was the filter, the turret and the pore helix were fairly well conserved, and the outer and the inner transmembrane domains of the protein were the most variable. Two of the new predicted channels were shown to be functional K + channels.

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          A potassium channel protein encoded by chlorella virus PBCV-1.

          The large chlorella virus PBCV-1, which contains double-stranded DNA (dsDNA), encodes a 94-codon open reading frame (ORF) that contains a motif resembling the signature sequence of the pore domain of potassium channel proteins. Phylogenetic analyses of the encoded protein, Kcv, indicate a previously unidentified type of potassium channel. The messenger RNA encoded by the ORF leads to functional expression of a potassium-selective conductance in Xenopus laevis oocytes. The channel blockers amantadine and barium, but not cesium, inhibit this conductance, in addition to virus plaque formation. Thus, PBCV-1 encodes the first known viral protein that functions as a potassium-selective channel and is essential in the virus life cycle.
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            Towards defining the chloroviruses: a genomic journey through a genus of large DNA viruses

            Background Giant viruses in the genus Chlorovirus (family Phycodnaviridae) infect eukaryotic green microalgae. The prototype member of the genus, Paramecium bursaria chlorella virus 1, was sequenced more than 15 years ago, and to date there are only 6 fully sequenced chloroviruses in public databases. Presented here are the draft genome sequences of 35 additional chloroviruses (287 – 348 Kb/319 – 381 predicted protein encoding genes) collected across the globe; they infect one of three different green algal species. These new data allowed us to analyze the genomic landscape of 41 chloroviruses, which revealed some remarkable features about these viruses. Results Genome colinearity, nucleotide conservation and phylogenetic affinity were limited to chloroviruses infecting the same host, confirming the validity of the three previously known subgenera. Clues for the existence of a fourth new subgenus indicate that the boundaries of chlorovirus diversity are not completely determined. Comparison of the chlorovirus phylogeny with that of the algal hosts indicates that chloroviruses have changed hosts in their evolutionary history. Reconstruction of the ancestral genome suggests that the last common chlorovirus ancestor had a slightly more diverse protein repertoire than modern chloroviruses. However, more than half of the defined chlorovirus gene families have a potential recent origin (after Chlorovirus divergence), among which a portion shows compositional evidence for horizontal gene transfer. Only a few of the putative acquired proteins had close homologs in databases raising the question of the true donor organism(s). Phylogenomic analysis identified only seven proteins whose genes were potentially exchanged between the algal host and the chloroviruses. Conclusion The present evaluation of the genomic evolution pattern suggests that chloroviruses differ from that described in the related Poxviridae and Mimiviridae. Our study shows that the fixation of algal host genes has been anecdotal in the evolutionary history of chloroviruses. We finally discuss the incongruence between compositional evidence of horizontal gene transfer and lack of close relative sequences in the databases, which suggests that the recently acquired genes originate from a still largely un-sequenced reservoir of genomes, possibly other unknown viruses that infect the same hosts.
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              An icosahedral algal virus has a complex unique vertex decorated by a spike.

              Paramecium bursaria Chlorella virus-1 is an icosahedrally shaped, 1,900-A-diameter virus that infects unicellular eukaryotic green algae. A 5-fold symmetric, 3D reconstruction using cryoelectron microscopy images has now shown that the quasiicosahedral virus has a unique vertex, with a pocket on the inside and a spike structure on the outside of the capsid. The pocket might contain enzymes for use in the initial stages of infection. The unique vertex consists of virally coded proteins, some of which have been identified. Comparison of shape, size, and location of the spike with similar features in bacteriophages T4 and P22 suggests that the spike might be a cell-puncturing device. Similar asymmetric features may have been missed in previous analyses of many other viruses that had been assumed to be perfectly icosahedral.
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                Author and article information

                Journal
                Viruses
                Viruses
                viruses
                Viruses
                MDPI
                1999-4915
                23 June 2020
                June 2020
                : 12
                : 6
                : 678
                Affiliations
                [1 ]School of Biological Sciences—Microbiology Program, University of Nebraska-Lincoln, Lincoln, NE 68588-0118, USA; murry.carter12@ 123456gmail.com
                [2 ]Nebraska Center for Virology, University of Nebraska-Lincoln, Lincoln, NE 68583-0900, USA; irina@ 123456unl.edu (I.V.A.); jghosh2@ 123456unl.edu (J.S.G.); roger.carlson@ 123456unl.edu (R.M.C.)
                [3 ]Department of Plant Pathology, University of Nebraska-Lincoln, Lincoln, NE 68583-0833, USA
                [4 ]Department of Chemistry and Biochemistry, Benedictine College, Atchison, KS 66002, USA; f.fitzgerald559@ 123456gmail.com
                [5 ]Membrane Biophysics, Department of Biology, Technische Universitat, 64287 Darmstadt, Germany; hertelb@ 123456bio.tu-darmstadt.de (B.H.); kukovetz@ 123456bio.tu-darmstadt.de (K.K.); rauh@ 123456bio.tu-darmstadt.de (O.R.); thiel@ 123456bio.tu-darmstadt.de (G.T.)
                Author notes
                [* ]Correspondence: jvanetten1@ 123456unl.edu ; Tel.: +1-402-472-3168
                Author information
                https://orcid.org/0000-0002-5063-0049
                Article
                viruses-12-00678
                10.3390/v12060678
                7354518
                32585987
                c2c541a2-4861-4f96-bf5e-6ff5ba426f75
                © 2020 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 03 June 2020
                : 19 June 2020
                Categories
                Article

                Microbiology & Virology
                chloroviruses,potassium ion channels,kcv channels,algal viruses
                Microbiology & Virology
                chloroviruses, potassium ion channels, kcv channels, algal viruses

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