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- Title
Probing the physiological role of the plastid outer-envelope membrane using the oemiR plasmid collection.
- Authors
Schwenkert, Serena; Wing Tung Lo; Szulc, Beata; Chun Kwan Yip; Pratt, Anna I.; Cusack, Siobhan A.; Brandt, Benjamin; Leister, Dario; Kunz, Hans-Henning
- Abstract
Plastids are the site of complex biochemical pathways, most prominently photosynthesis. The organelle evolved through endosymbiosis with a cyanobacterium, which is exemplified by the outer envelope membrane that harbors more than 40 proteins in Arabidopsis. Their evolutionary conservation indicates high significance for plant cell function. While a few proteins are well-studied as part of the protein translocon complex the majority of outer envelope protein functions is unclear. Gaining a deeper functional understanding has been complicated by the lack of observable loss-of-function mutant phenotypes, which is often rooted in functional genetic redundancy. Therefore, we designed outer envelope-specific artificial micro RNAs (oemiRs) capable of downregulating transcripts from several loci simultaneously. We successfully tested oemiR function by performing a proof-of-concept screen for pale and cold-sensitive mutants. An in-depth analysis of pale mutant alleles deficient in the translocon component TOC75 using proteomics provided new insights into putative compensatory import pathways. The cold stress screen not only recapitulated 3 previously known phenotypes of cold-sensitive mutants but also identified 4 mutants of additional oemiR outer envelope loci. Altogether our study revealed a role of the outer envelope to tolerate cold conditions and showcasts the power of the oemiR collection to research the significance of outer envelope proteins.
- Subjects
MICRORNA; ARABIDOPSIS proteins; CHLOROPLAST membranes; CELL physiology; PLASTIDS; PROTEOMICS
- Publication
G3: Genes | Genomes | Genetics, 2023, Vol 13, Issue 10, p1
- ISSN
2160-1836
- Publication type
Article
- DOI
10.1093/g3journal/jkad187