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      Telomerase RNAs in land plants

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          Abstract

          To elucidate the molecular nature of evolutionary changes of telomeres in the plant order Asparagales, we aimed to characterize telomerase RNA subunits (TRs) in these plants. The unusually long telomere repeat unit in Allium plants (12 nt) allowed us to identify TRs in transcriptomic data of representative species of the Allium genus. Orthologous TRs were then identified in Asparagales plants harbouring telomere DNA composed of TTAGGG (human type) or TTTAGGG ( Arabidopsis-type) repeats. Further, we identified TRs across the land plant phylogeny, including common model plants, crop plants, and plants with unusual telomeres. Several lines of functional testing demonstrate the templating telomerase function of the identified TRs and disprove a functionality of the only previously reported plant telomerase RNA in Arabidopsis thaliana. Importantly, our results change the existing paradigm in plant telomere biology which has been based on the existence of a relatively conserved telomerase reverse transcriptase subunit (TERT) associating with highly divergent TRs even between closely related plant taxa. The finding of a monophyletic origin of genuine TRs across land plants opens the possibility to identify TRs directly in transcriptomic or genomic data and/or predict telomere sequences synthesized according to the respective TR template region.

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          Most cited references57

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          Genome-wide insertional mutagenesis of Arabidopsis thaliana.

          J Alonso (2003)
          Over 225,000 independent Agrobacterium transferred DNA (T-DNA) insertion events in the genome of the reference plant Arabidopsis thaliana have been created that represent near saturation of the gene space. The precise locations were determined for more than 88,000 T-DNA insertions, which resulted in the identification of mutations in more than 21,700 of the approximately 29,454 predicted Arabidopsis genes. Genome-wide analysis of the distribution of integration events revealed the existence of a large integration site bias at both the chromosome and gene levels. Insertion mutations were identified in genes that are regulated in response to the plant hormone ethylene.
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            Identification of a specific telomere terminal transferase activity in Tetrahymena extracts.

            We have found a novel activity in Tetrahymena cell free extracts that adds tandem TTGGGG repeats onto synthetic telomere primers. The single-stranded DNA oligonucleotides (TTGGGG)4 and TGTGTGGGTGTGTGGGTGTGTGGG, consisting of the Tetrahymena and yeast telomeric sequences respectively, each functioned as primers for elongation, while (CCCCAA)4 and two nontelomeric sequence DNA oligomers did not. Efficient synthesis of the TTGGGG repeats depended only on addition of micromolar concentrations of oligomer primer, dGTP, and dTTP to the extract. The activity was sensitive to heat and proteinase K treatment. The repeat addition was independent of both endogenous Tetrahymena DNA and the endogenous alpha-type DNA polymerase; and a greater elongation activity was present during macronuclear development, when a large number of telomeres are formed and replicated, than during vegetative cell growth. We propose that the novel telomere terminal transferase is involved in the addition of telomeric repeats necessary for the replication of chromosome ends in eukaryotes.
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              A telomeric sequence in the RNA of Tetrahymena telomerase required for telomere repeat synthesis.

              The telomerase enzyme of Tetrahymena synthesizes repeats of the telomeric DNA sequence TTGGGG de novo in the absence of added template. The essential RNA component of this ribonucleoprotein enzyme has now been cloned and found to contain the sequence CAACCCCAA, which seems to be the template for the synthesis of TTGGGG repeats.
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                Author and article information

                Journal
                Nucleic Acids Res
                Nucleic Acids Res
                nar
                Nucleic Acids Research
                Oxford University Press
                0305-1048
                1362-4962
                10 October 2019
                08 August 2019
                08 August 2019
                : 47
                : 18
                : 9842-9856
                Affiliations
                [1 ] Department of Cell Biology and Radiobiology, Institute of Biophysics of the Czech Academy of Sciences , v.v.i., Brno CZ-61265, Czech Republic
                [2 ] Laboratory of Functional Genomics and Proteomics, NCBR, Faculty of Science, Masaryk University , Brno CZ-61137, Czech Republic
                [3 ] Mendel Centre for Plant Genomics and Proteomics, CEITEC, Masaryk University , Brno CZ-62500, Czech Republic
                [4 ] Max Perutz Labs, University of Vienna , Dr. Bohr Gasse 9, A-1030, Vienna, Austria
                Author notes
                To whom correspondence should be addressed. Tel: +420 549494003; Fax: +420 549492654; Email: fajkus@ 123456sci.muni.cz
                Correspondence may also be addressed to Eva Sýkorová. Tel: +420 541517199; Email: evin@ 123456ibp.cz

                The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors.

                Author information
                http://orcid.org/0000-0002-3112-1716
                Article
                gkz695
                10.1093/nar/gkz695
                6765143
                31392988
                ac284ddc-3eaa-4633-9ace-fdb42adac87a
                © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 30 July 2019
                : 26 July 2019
                : 11 July 2019
                Page count
                Pages: 15
                Funding
                Funded by: Czech Science Foundation 10.13039/501100001824
                Award ID: 17-09644S
                Funded by: ERDF 10.13039/501100008530
                Award ID: CZ.02.1.01/0.0/0.0/15_003/0000477
                Funded by: Ministry of Education, Youth and Sports 10.13039/501100001823
                Award ID: LQ1601
                Categories
                RNA and RNA-protein complexes

                Genetics
                Genetics

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