ultrastructural and molecular analyses reveal enhanced nucleolar activity in medicago truncatula cells overexpressing the mttdp2α gene

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ID: 137725
2018
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Abstract
The role of tyrosyl-DNA phosphodiesterase 2 (Tdp2) involved in the repair of 5′-end-blocking DNA lesions is still poorly explored in plants. To gain novel insights, Medicago truncatula suspension cultures overexpressing the MtTdp2α gene (Tdp2α-13C and Tdp2α-28 lines, respectively) and a control (CTRL) line carrying the empty vector were investigated. Transmission electron microscopy (TEM) revealed enlarged nucleoli (up to 44% expansion of the area, compared to CTRL), the presence of nucleolar vacuoles, increased frequency of multinucleolate cells (up to 4.3-fold compared to CTRL) and reduced number of ring-shaped nucleoli in Tdp2α-13C and Tdp2α-28 lines. Ultrastructural data suggesting for enhanced nucleolar activity in MtTdp2α-overexpressing lines were integrated with results from bromouridine incorporation. The latter revealed an increase of labeled transcripts in both Tdp2α-13C and Tdp2α-28 cells, within the nucleolus and in the extra-nucleolar region. MtTdp2α-overexpressing cells showed tolerance to etoposide, a selective inhibitor of DNA topoisomerase II, as evidenced by DNA diffusion assay. TEM analysis revealed etoposide-induced rearrangements within the nucleolus, resembling the nucleolar caps observed in animal cells under transcription impairment. Based on these findings it is evident that MtTdp2α-overexpression enhances nucleolar activity in plant cells.
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macovei2018frontiersultrastructural Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Anca Macovei, Matteo Faè, Marco Biggiogera, Susana de Sousa Araújo, Daniela Carbonera, Alma Balestrazzi
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Journal phytochemistry letters
Year 2018
DOI
10.3389/fpls.2018.00596
URL
http://journal.frontiersin.org/article/10.3389/fpls.2018.00596/full
Keywords
transmission electron microscopy medicago truncatula

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