Introduction of a Heterologous Editing Site into the Tobacco Plastid Genome: the Lack of RNA Editing Leads to a Mutant Phenotype

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 1994 Oct 3

PMID 7925303

Citations 80

Authors

R Bock

H Kossel

P Maliga

Affiliations

Soon will be listed here.

Abstract

The psbF mRNA is edited in spinach plastids by a C to U conversion, changing a serine to a conserved phenylalanine codon. In tobacco at this position a phenylalanine codon is present at the DNA level, and the psbF mRNA here is not edited. To test if the psbF editing capacity is evolutionarily conserved, the tobacco psbF gene was modified to match the corresponding spinach sequence. The endogenous tobacco gene was replaced with the modified copy using biolistic transformation. We report here that the heterologous editing site remains unmodified in transplastomic tobacco plants. The lack of editing is associated with slower growth, lowered chlorophyll content and high chlorophyll fluorescence, a phenotype characteristic of photosynthetic mutants. This finding confirms that the editing of the psbF mRNA is an essential processing step for protein function and thus provides direct proof for the biological significance of plant organellar RNA editing. Given that a mutant phenotype is associated with the lack of editing, it seems likely that the evolutionary loss of the site-specific capacity for psbF editing was preceded by the mutation that eliminated the editing requirement.

Citing Articles

Membrane protein provision controls prothylakoid biogenesis in tobacco etioplasts.

Li B, Armarego-Marriott T, Kowalewska L, Thiele W, Erban A, Ruf S Plant Cell. 2024; .

PMID: 39321213 PMC: 11638105. DOI: 10.1093/plcell/koae259.

Transgene insertion into the plastid genome alters expression of adjacent native chloroplast genes at the transcriptional and translational levels.

Ghandour R, Gao Y, Laskowski J, Barahimipour R, Ruf S, Bock R Plant Biotechnol J. 2022; 21(4):711-725.

PMID: 36529916 PMC: 10037153. DOI: 10.1111/pbi.13985.

Genome-Wide Identification and Characterization of RNA/DNA Differences Associated with Infection in Wheat.

Yang G, Pan Y, Zhao Q, Huang J, Pan W, Cui L Int J Mol Sci. 2022; 23(14).

PMID: 35887327 PMC: 9316857. DOI: 10.3390/ijms23147982.

Plastid Transformation in Tomato: A Vegetable Crop and Model Species.

Ruf S, Bock R Methods Mol Biol. 2021; 2317:217-228.

PMID: 34028771 DOI: 10.1007/978-1-0716-1472-3_11.

Advancing organelle genome transformation and editing for crop improvement.

Li S, Chang L, Zhang J Plant Commun. 2021; 2(2):100141.

PMID: 33898977 PMC: 8060728. DOI: 10.1016/j.xplc.2021.100141.

References

Covello P, Gray M . RNA editing in plant mitochondria. Nature. 1989; 341(6243):662-6. DOI: 10.1038/341662a0. View

Hiesel R, Wissinger B, Schuster W, Brennicke A . RNA editing in plant mitochondria. Science. 1989; 246(4937):1632-4. DOI: 10.1126/science.2480644. View

Zeltz P, Hess W, Neckermann K, Borner T, Kossel H . Editing of the chloroplast rpoB transcript is independent of chloroplast translation and shows different patterns in barley and maize. EMBO J. 1993; 12(11):4291-6. PMC: 413725. DOI: 10.1002/j.1460-2075.1993.tb06113.x. View

Svab Z, Hajdukiewicz P, Maliga P . Stable transformation of plastids in higher plants. Proc Natl Acad Sci U S A. 1990; 87(21):8526-30. PMC: 54989. DOI: 10.1073/pnas.87.21.8526. View

Maier R, Neckermann K, Hoch B, Akhmedov N, Kossel H . Identification of editing positions in the ndhB transcript from maize chloroplasts reveals sequence similarities between editing sites of chloroplasts and plant mitochondria. Nucleic Acids Res. 1992; 20(23):6189-94. PMC: 334503. DOI: 10.1093/nar/20.23.6189. View

Kudla J, Igloi G, Metzlaff M, Hagemann R, Kossel H . RNA editing in tobacco chloroplasts leads to the formation of a translatable psbL mRNA by a C to U substitution within the initiation codon. EMBO J. 1992; 11(3):1099-103. PMC: 556551. DOI: 10.1002/j.1460-2075.1992.tb05149.x. View

Kanevski I, Maliga P . Relocation of the plastid rbcL gene to the nucleus yields functional ribulose-1,5-bisphosphate carboxylase in tobacco chloroplasts. Proc Natl Acad Sci U S A. 1994; 91(5):1969-73. PMC: 43286. DOI: 10.1073/pnas.91.5.1969. View

Gray M, Covello P . RNA editing in plant mitochondria and chloroplasts. FASEB J. 1993; 7(1):64-71. DOI: 10.1096/fasebj.7.1.8422976. View

Svab Z, Maliga P . High-frequency plastid transformation in tobacco by selection for a chimeric aadA gene. Proc Natl Acad Sci U S A. 1993; 90(3):913-7. PMC: 45780. DOI: 10.1073/pnas.90.3.913. View

10.

Bachmann B, Luke W, Hunsmann G . Improvement of PCR amplified DNA sequencing with the aid of detergents. Nucleic Acids Res. 1990; 18(5):1309. PMC: 330471. DOI: 10.1093/nar/18.5.1309. View

11.

Gualberto J, Lamattina L, Bonnard G, Weil J, Grienenberger J . RNA editing in wheat mitochondria results in the conservation of protein sequences. Nature. 1989; 341(6243):660-2. DOI: 10.1038/341660a0. View

12.

Shinozaki K, Ohme M, Tanaka M, Wakasugi T, Hayashida N, Matsubayashi T . The complete nucleotide sequence of the tobacco chloroplast genome: its gene organization and expression. EMBO J. 1986; 5(9):2043-2049. PMC: 1167080. DOI: 10.1002/j.1460-2075.1986.tb04464.x. View

13.

Carrillo N, Seyer P, Tyagi A, Herrmann R . Cytochrome b-559 genes from Oenothera hookeri and Nicotiana tabacum show a remarkably high degree of conservation as compared to spinach. The enigma of cytochrome b-559: highly conserved genes and proteins but no known function. Curr Genet. 1986; 10(8):619-24. DOI: 10.1007/BF00418129. View

14.

Hoch B, Maier R, Appel K, Igloi G, Kossel H . Editing of a chloroplast mRNA by creation of an initiation codon. Nature. 1991; 353(6340):178-80. DOI: 10.1038/353178a0. View

15.

Wissinger B, Schuster W, Brennicke A . Species-specific RNA editing patterns in the mitochondrial rps13 transcripts of Oenothera and Daucus. Mol Gen Genet. 1990; 224(3):389-95. DOI: 10.1007/BF00262433. View

16.

Covello P, Gray M . Differences in editing at homologous sites in messenger RNAs from angiosperm mitochondria. Nucleic Acids Res. 1990; 18(17):5189-96. PMC: 332141. DOI: 10.1093/nar/18.17.5189. View

17.

Pakrasi H, De Ciechi P, Whitmarsh J . Site directed mutagenesis of the heme axial ligands of cytochrome b559 affects the stability of the photosystem II complex. EMBO J. 1991; 10(7):1619-27. PMC: 452830. DOI: 10.1002/j.1460-2075.1991.tb07684.x. View

18.

Bock R, Hagemann R, Kossel H, Kudla J . Tissue- and stage-specific modulation of RNA editing of the psbF and psbL transcript from spinach plastids--a new regulatory mechanism?. Mol Gen Genet. 1993; 240(2):238-44. DOI: 10.1007/BF00277062. View

19.

Iwabuchi M, Kyozuka J, Shimamoto K . Processing followed by complete editing of an altered mitochondrial atp6 RNA restores fertility of cytoplasmic male sterile rice. EMBO J. 1993; 12(4):1437-46. PMC: 413355. DOI: 10.1002/j.1460-2075.1993.tb05787.x. View