Overexpression of Active or Inactive RNase J Disrupts Chloroplast RNA Quality Control in Arabidopsis
Year: 2026
Type: preprint
Abstract: SUMMARY The chloroplast is a semi-autonomous organelle with a compact genome encoding ∼100 expressed genes. Numerous and widespread promoters coupled with inefficient transcription termination leads to a highly complex primary transcriptome that undergoes extensive RNA quality control orchestrated by RNA binding proteins and ribonucleases. Among the ribonucleases, only RNase J (RNJ) is absolutely required for plant germination and survival, likely because its deficiency leads to massive accumulation of antisense RNA which inhibits translation. As one approach to understanding the selectivity of RNJ for antisense RNA and noncoding regions, we created Arabidopsis lines overexpressing catalytically inactive versions of the enzyme designed to trap its substrates. Initially, transgenic plants appeared phenotypically normal. However, crosses used to generate homozygous transgenic lines yielded progeny that developed varying degrees of chlorosis within 2-3 weeks. Concomitantly, these plants exhibited chloroplast transcriptome alterations similar to RNJ-silenced lines, along with mRNA changes consistent with retrograde signaling. This chlorosis could be correlated to the likely decay of RNJ protein, but not its endogenous transcript. The result could be replicated by attempting to overexpress a wild-type version of RNJ, in which case even one transgene copy was sufficient to induce chlorosis. Our results suggest that RNJ is under protein expression level surveillance to prevent wholesale destruction of the plastid transcriptome.
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
Funder: Institut Universitaire de France
OpenAlex ID: W7220671659