Assessment of Protein Synthesis in Mitochondria Isolated from Rosette Leaves and Liquid Culture Seedlings of Arabidopsis

Kwasniak-Owczarek, Malgorzata; Tomal, Artur; Jańska, Hanna

doi:10.1007/978-1-0716-1653-6_14

Cited by 5 publications

(2 citation statements)

References 21 publications

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“…Sodium acetate, a substrate only for bacterial translation, was used to evaluate bacterial contamination during mitochondrial purification. We could identify several distinct protein bands, including Atp1, Cob, Cox2, and Atp9, based on their molecular mass ( 34 , 35 ). Atp1, Cob, Cox2, and Atp9 had a slower rate of translation in the mtran double-knockout mutants than in the WT.…”

Section: Mtran1 and Mtran2 Are Required For Translation In Mitochondriamentioning

confidence: 99%

An mTRAN-mRNA interaction mediates mitochondrial translation initiation in plants

Tran,

Schmitt,

Lama

et al. 2023

Science

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Plant mitochondria represent the largest group of respiring organelles on the planet. Plant mitochondrial messenger RNAs (mRNAs) lack Shine-Dalgarno-like ribosome-binding sites, so it is unknown how plant mitoribosomes recognize mRNA. We show that “mitochondrial translation factors” mTRAN1 and mTRAN2 are land plant–specific proteins, required for normal mitochondrial respiration chain biogenesis. Our studies suggest that mTRANs are noncanonical pentatricopeptide repeat (PPR)–like RNA binding proteins of the mitoribosomal “small” subunit. We identified conserved Adenosine (A)/Uridine (U)-rich motifs in the 5′ regions of plant mitochondrial mRNAs. mTRAN1 binds this motif, suggesting that it is a mitoribosome homing factor to identify mRNAs. We demonstrate that mTRANs are likely required for translation of all plant mitochondrial mRNAs. Plant mitochondrial translation initiation thus appears to use a protein-mRNA interaction that is divergent from bacteria or mammalian mitochondria.

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Section: Mtran1 and Mtran2 Are Required For Translation In Mitochondriamentioning

confidence: 99%

An mTRAN-mRNA interaction mediates mitochondrial translation initiation in plants

Tran,

Schmitt,

Lama

et al. 2023

Science

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show abstract

“…An in-organelle protein synthesis assay was performed following a procedure as described (Kwasniak-Owczarek et al, 2022). A translation mix (5 mM KH 2 PO 4 , 2 mM guanosine 5'-triphosphate, 0.4 M mannitol, 60 mM KCl, 2 mM DTT, 50 mM HEPES, 10 mM MgCl 2 , pH 7.0) was divided into two equal volumes for mitochondrial translation and bacterial control.…”

Section: Analysis Of In Organelle Protein Synthesismentioning

confidence: 99%

Protein aggregation in plant mitochondria inhibits translation and induces an NAC017-dependent ethylene-associated unfolded protein response

Song

Hou

et al. 2023

Preprint

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Loss of Lon1 in plant mitochondria led to stunted plant growth and accumulation of nuclear-encoded mitochondrial proteins, including Lon1 substrates, while mitochondrial-encoded proteins typically decreased in abundance. Lon1 mutants contained protein aggregates in the mitochondria matrix which were enriched in PPR-containing proteins and ribosomal subunits of the translation apparatus and were slowed in mitochondrial RNA splicing, editing and general translation rate. Transcriptome analysis showed multiple organellar unfolded protein responses involving ethylene biosynthesis were induced by either Lon1 loss, mitochondrial ribosomal protein loss, translation or respiratory inhibition and most were regulated by the mitochondrial retrograde signaling pathway dependent on the transcription factor NAC017. The short hypocotyl in lon1 mutants during skotomorphogenesis was partially rescued by ethylene inhibitors and mutants showed higher ethylene production rates than wildtype. Together this provides multiple steps in the link between loss of Lon1 and its whole plant phenotype.

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Purification of Leaf Mitochondria from Arabidopsis thaliana Using Percoll Density Gradients

Tran

Aken

2021

Methods in Molecular Biology

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Assessment of Protein Synthesis in Mitochondria Isolated from Rosette Leaves and Liquid Culture Seedlings of Arabidopsis

Cited by 5 publications

References 21 publications

An mTRAN-mRNA interaction mediates mitochondrial translation initiation in plants

An mTRAN-mRNA interaction mediates mitochondrial translation initiation in plants

Protein aggregation in plant mitochondria inhibits translation and induces an NAC017-dependent ethylene-associated unfolded protein response

Purification of Leaf Mitochondria from Arabidopsis thaliana Using Percoll Density Gradients

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