Cell cycle stage-specific differential expression of topoisomerase I in tobacco BY-2 cells and its ectopic overexpression and knockdown unravels its crucial role in plant morphogenesis and development

Plant Sci. 2015 Nov:240:182-92. doi: 10.1016/j.plantsci.2015.09.016. Epub 2015 Sep 25.

Abstract

DNA topoisomerases catalyze the inter-conversion of different topological forms of DNA. Cell cycle coupled differential accumulation of topoisomerase I (Topo I) revealed biphasic expression maximum at S-phase and M/G1-phase of cultured synchronized tobacco BY-2 cells. This suggested its active role in resolving topological constrains during DNA replication (S-phase) and chromosome decondensation (M/G1 phase). Immuno-localization revealed high concentrations of Topo I in nucleolus. Propidium iodide staining and Br-UTP incorporation patterns revealed direct correlation between immunofluorescence intensity and rRNA transcription activity within nucleolus. Immuno-stained chromosomes during metaphase and anaphase suggested possible role of Topo I in resolving topological constrains during mitotic chromosome condensation. Inhibitor studies showed that in comparison to Topo I, Topo II was essential in resolving topological constrains during chromosome condensation. Probably, Topo II substituted Topo I functioning to certain extent during chromosome condensation, but not vice-versa. Transgenic Topo I tobacco lines revealed morphological abnormalities and highlighted its crucial role in plant morphogenesis and development.

Keywords: Chromosome condensation; Nicotiana tabaccum; Plant development; Synchronized BY-2 cell; Topoisomerase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle
  • Cells, Cultured
  • DNA Topoisomerases, Type I / genetics*
  • DNA Topoisomerases, Type I / metabolism
  • Ectopic Gene Expression
  • Fluorescent Antibody Technique
  • Gene Expression Regulation, Plant*
  • Gene Knockdown Techniques
  • Nicotiana / genetics*
  • Nicotiana / growth & development
  • Nicotiana / metabolism
  • Plant Proteins / genetics*
  • Plant Proteins / metabolism
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / growth & development
  • Plants, Genetically Modified / metabolism

Substances

  • Plant Proteins
  • DNA Topoisomerases, Type I