Import of a Precursor Protein into Chloroplasts is Inhibited by the Herbicide Glyphosate

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 1988 May 1

PMID 16453841

Citations 22

Authors

G Della-Cioppa

G M Kishore

Affiliations

Soon will be listed here.

Abstract

Import of the precursor to 5-enolpyruvylshikimate-3-phosphate synthase (pEPSPS) into chloroplasts is inhibited by the herbicide glyphosate. Inhibition of import is maximal at glyphosate concentrations of >/=10 mum and occurs only when pEPSPS is present as a ternary complex of enzyme-shikimate-3-phosphate-glyphosate. Glyphosate alone had no effect on the import of pEPSPS since it is not known to interact with the enzyme in the absence of shikimate-3-phosphate. Experiments with wild-type and glyphosate-resistant mutant forms of pEPSPS show that inhibition of import is directly proportional to the binding constants for glyphosate. Inhibition of import is thus a direct consequence of glyphosate binding to the enzyme-shikimate-3-phosphate complex. The potential for non-specific effects of glyphosate on the chloroplast transport mechanism has been discounted by showing that import of another chloroplast-designated protein was unaffected by high concentrations of glyphosate and shikimate-3-phosphate. The mechanism of import inhibition by glyphosate is consistent with a precursor unfolding/refolding model.

Citing Articles

Glyphosate: Uses Other Than in Glyphosate-Resistant Crops, Mode of Action, Degradation in Plants, and Effects on Non-target Plants and Agricultural Microbes.

Duke S Rev Environ Contam Toxicol. 2021; 255:1-65.

PMID: 33895876 DOI: 10.1007/398_2020_53.

Ecotoxicological Studies on the Effect of Roundup (Glyphosate Formulation) on Marine Benthic Microalgae.

Sylwestrzak Z, Zgrundo A, Pniewski F Int J Environ Res Public Health. 2021; 18(3).

PMID: 33498564 PMC: 7908156. DOI: 10.3390/ijerph18030884.

Molecular cloning and structural analysis of the phosphate translocator from pea chloroplasts and its comparison to the spinach phosphate translocator.

Willey D, Fischer K, Wachter E, Link T, Flugge U Planta. 2013; 183(3):451-61.

PMID: 24193755 DOI: 10.1007/BF00197745.

Stability and expression of amplified EPSPS genes in glyphosate resistant tobacco cells and plantlets.

Jones J, Goldsbrough P, Weller S Plant Cell Rep. 2013; 15(6):431-6.

PMID: 24178424 DOI: 10.1007/BF00232070.

Purification and properties of a glyphosate-tolerant 5-enolpyruvylshikimate 3-phosphate synthase from the cyanobacterium Anabaena variabilis.

Powell H, Kerby N, Rowell P, Mousdale D, Coggins J Planta. 2013; 188(4):484-90.

PMID: 24178379 DOI: 10.1007/BF00197039.

References

Chua N, Schmidt G . Post-translational transport into intact chloroplasts of a precursor to the small subunit of ribulose-1,5-bisphosphate carboxylase. Proc Natl Acad Sci U S A. 1978; 75(12):6110-4. PMC: 393128. DOI: 10.1073/pnas.75.12.6110. View

Pain D, Blobel G . Protein import into chloroplasts requires a chloroplast ATPase. Proc Natl Acad Sci U S A. 1987; 84(10):3288-92. PMC: 304854. DOI: 10.1073/pnas.84.10.3288. View

Eilers M, Schatz G . Binding of a specific ligand inhibits import of a purified precursor protein into mitochondria. Nature. 1986; 322(6076):228-32. DOI: 10.1038/322228a0. View

Chen W, DOUGLAS M . Phosphodiester bond cleavage outside mitochondria is required for the completion of protein import into the mitochondrial matrix. Cell. 1987; 49(5):651-8. DOI: 10.1016/0092-8674(87)90541-1. View

Randall L, Hardy S . Correlation of competence for export with lack of tertiary structure of the mature species: a study in vivo of maltose-binding protein in E. coli. Cell. 1986; 46(6):921-8. DOI: 10.1016/0092-8674(86)90074-7. View

Flugge U, Hinz G . Energy dependence of protein translocation into chloroplasts. Eur J Biochem. 1986; 160(3):563-70. DOI: 10.1111/j.1432-1033.1986.tb10075.x. View

Boocock M, Coggins J . Kinetics of 5-enolpyruvylshikimate-3-phosphate synthase inhibition by glyphosate. FEBS Lett. 1983; 154(1):127-33. DOI: 10.1016/0014-5793(83)80888-6. View

Della-Cioppa G, Bauer S, Klein B, Shah D, Fraley R, Kishore G . Translocation of the precursor of 5-enolpyruvylshikimate-3-phosphate synthase into chloroplasts of higher plants in vitro. Proc Natl Acad Sci U S A. 1986; 83(18):6873-7. PMC: 386612. DOI: 10.1073/pnas.83.18.6873. View

Shah D, Horsch R, Klee H, Kishore G, WINTER J, Tumer N . Engineering herbicide tolerance in transgenic plants. Science. 1986; 233(4762):478-81. DOI: 10.1126/science.233.4762.478. View

10.

Lubben T, Bansberg J, Keegstra K . Stop-transfer regions do not halt translocation of proteins into chloroplasts. Science. 1987; 238(4830):1112-4. DOI: 10.1126/science.238.4830.1112. View

11.

Schleyer M, Neupert W . Transport of proteins into mitochondria: translocational intermediates spanning contact sites between outer and inner membranes. Cell. 1985; 43(1):339-50. DOI: 10.1016/0092-8674(85)90039-x. View

12.

Schmidt G, Mishkind M . The transport of proteins into chloroplasts. Annu Rev Biochem. 1986; 55:879-912. DOI: 10.1146/annurev.bi.55.070186.004311. View

13.

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

14.

Steinrucken H, Amrhein N . The herbicide glyphosate is a potent inhibitor of 5-enolpyruvyl-shikimic acid-3-phosphate synthase. Biochem Biophys Res Commun. 1980; 94(4):1207-12. DOI: 10.1016/0006-291x(80)90547-1. View