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Distinct thresholds condition sense posttranscriptional gene silencing initiation and amplification.

Martin Lacroix1,2, Nicolas Butel1, Andana Barrios1

  • 1Université Paris-Saclay, INRAE, AgroParisTech, Institut Jean-Pierre Bourgin for Plant Sciences, Versailles 78000, France.

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Gene expression levels dictate the amplification of sense post-transcriptional gene silencing (S-PTGS). Distinct RNA thresholds control S-PTGS initiation and systemic spread, even in endogenous genes.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Plant Science

Background:

  • Pro35S-driven sense transgenes frequently undergo post-transcriptional gene silencing (S-PTGS).
  • The precise conditions governing S-PTGS initiation and systemic amplification remain largely unknown.
  • Understanding these mechanisms is crucial for transgene stability and gene regulation studies.

Purpose of the Study:

  • To elucidate the factors controlling S-PTGS initiation and amplification.
  • To investigate the relationship between transgene expression levels and S-PTGS capacity.
  • To determine the thresholds required for S-PTGS induction and systemic spread.

Main Methods:

  • Genetic screens utilizing chromatin-related mutations.
  • Analysis of independent transgenic plant lines with varying transgene expression.
  • Experimental setup combining inducible/tissue-specific transgenes with S-PTGS-prone loci.

Main Results:

  • Chromatin mutations modulating transgene expression affected S-PTGS amplification but not initiation.
  • A direct correlation was observed between transgene expression levels and S-PTGS amplification capacity.
  • Transient expression exceeding a threshold was sufficient to initiate systemic S-PTGS.
  • Distinct thresholds for aberrant RNA (initiation) and target mRNA (amplification) were proposed.

Conclusions:

  • Transgene expression levels are critical for S-PTGS amplification.
  • Separate thresholds govern S-PTGS initiation and systemic spread.
  • The proposed model applies to endogenous genes, with RNA Quality Control and DCL proteins acting as protective layers.