Abstract

Splicing of plant organellar transcripts is facilitated by members of a large protein family, the pentatricopeptide repeat proteins. We have identified a pentatricopeptide repeat protein in a genetic screen for mutants resistant to inhibition of root growth by buthionine sulfoximine (BSO), an inhibitor of glutathione synthesis and consequently named BIR6 (BSO-insensitive roots 6). BIR6 is involved in splicing of intron 1 of the mitochondrial nad7 transcript. Loss-of-function mutations in BIR6 result in a strongly reduced accumulation of fully processed nad7 transcript. This affects assembly of Complex I and results in moderate growth retardation. In agreement with disruption of Complex I function, the genes encoding alternative NADH oxidizing enzymes are induced in the mutant, and the mutant plants are less sensitive to mannitol and salt stress. Mutation in the BIR6 gene allowed normal root growth in presence of BSO and strongly attenuated depletion of glutathione content at these conditions. The same phenotype was observed with other mutants affected in function of Complex I, thus reinforcing the importance of Complex I function for cellular redox homeostasis.

Highlights

  • Splicing of plant organellar transcripts is facilitated by members of a large protein family, the pentatricopeptide repeat proteins

  • BIR6 Is Involved in Synthesis of the NAD7 Subunit of Complex

  • It is similar in structure to most other PPR proteins shown to be involved in RNA splicing, including OTP43 shown to be required for splicing of the first intron of Arabidopsis mitochondrial nad1 transcripts [36] and the maize chloroplast splicing factor PPR5 [43]

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Summary

Identification of a Pentatricopeptide Repeat Protein

We have identified a pentatricopeptide repeat protein in a genetic screen for mutants resistant to inhibition of root growth by buthionine sulfoximine (BSO), an inhibitor of glutathione synthesis and named BIR6 (BSO-insensitive roots 6). Loss-of-function mutations in BIR6 result in a strongly reduced accumulation of fully processed nad transcript. This affects assembly of Complex I and results in moderate growth retardation. We describe cloning of a mutant in a PPR protein isolated in a genetic screen to find mutants resistant to inhibition of root growth by BSO. We show that this protein is required. For efficient splicing of nad transcripts encoding a subunit of Complex I of the mitochondrial respiratory chain

EXPERIMENTAL PROCEDURES
RESULTS
Disruption of Complex I Affects
Findings
DISCUSSION
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