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PMID: 19635803 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Unraveling delta1-pyrroline-5-carboxylate-proline cycle in plants by uncoupled expression of proline oxidation enzymes.

The Journal of biological chemistry ·Vol. 284 ·No. 39 ·2009-09-25 ·Pages 26482-92

Miller G, Honig A, Stein H, Suzuki N, Mittler R, Zilberstein A

Abstract

The two-step oxidation of proline in all eukaryotes is performed at the inner mitochondrial membrane by the consecutive action of proline dehydrogenase (ProDH) that produces Delta(1)-pyrroline-5-carboxylate (P5C) and P5C dehydrogenase (P5CDH) that oxidizes P5C to glutamate. This catabolic route is down-regulated in plants during osmotic stress, allowing free Pro accumulation. We show here that overexpression of MsProDH in tobacco and Arabidopsis or impairment of P5C oxidation in the Arabidopsis p5cdh mutant did not change the cellular Pro to P5C ratio under ambient and osmotic stress conditions, indicating that P5C excess was reduced to Pro in a mitochondrial-cytosolic cycle. This cycle, involving ProDH and P5C reductase, exists in animal cells and now demonstrated in plants. As a part of the cycle, Pro oxidation by the ProDH-FAD complex delivers electrons to the electron transport chain. Hyperactivity of the cycle, e.g. when an excess of exogenous l-Pro is provided, generates mitochondrial reactive oxygen species (ROS) by delivering electrons to O(2), as demonstrated by the mitochondria-specific MitoSox staining of superoxide ions. Lack of P5CDH activity led to higher ROS production under dark and light conditions in the presence of Pro excess, as well as rendered plants hypersensitive to heat stress. Balancing mitochondrial ROS production during increased Pro oxidation is therefore critical for avoiding Pro-related toxic effects. Hence, normal oxidation of P5C to Glu by P5CDH is key to prevent P5C-Pro intensive cycling and avoid ROS production from electron run-off.

MeSH Terms
1-Pyrroline-5-Carboxylate Dehydrogenase/genetics,metabolism Arabidopsis/enzymology,genetics,metabolism Arabidopsis Proteins/genetics,metabolism Droughts Gene Expression Regulation, Enzymologic/drug effects Gene Expression Regulation, Plant/drug effects Glutamic Acid/metabolism Hot Temperature Medicago sativa/enzymology,genetics Mitochondria/drug effects,enzymology,metabolism Mutation Oxidation-Reduction Plants/enzymology,genetics,metabolism Plants, Genetically Modified Proline/metabolism,pharmacology Proline Oxidase/genetics,metabolism Pyrroles/metabolism Reactive Oxygen Species/metabolism Reverse Transcriptase Polymerase Chain Reaction Sodium Chloride/pharmacology Tobacco/enzymology,genetics,metabolism
Chemicals
Arabidopsis Proteins Pyrroles Reactive Oxygen Species delta-1-pyrroline-5-carboxylate Glutamic Acid Sodium Chloride Proline 1-Pyrroline-5-Carboxylate Dehydrogenase ALDH12A1 protein, Arabidopsis Proline Oxidase
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Miller Gad
Department of Plant Science, Tel Aviv University, Tel-Aviv 69978, Israel.
Honig Arik
Stein Hanan
Suzuki Nobuhiro
Mittler Ron
Zilberstein Aviah
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
1083-351X
DOI
10.1074/jbc.M109.009340
Published
2009-09-25
Epub
2009-00-27
Pages
26482-92
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC2785336
Subset
IM
External Links
PubMed source DOI: 10.1074/jbc.M109.009340 PMC Full Text View journal details
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