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Analysis of cytosolic and plastidic serine acetyltransferase mutants and subcellular metabolite distributions suggests interplay of the cellular compartments for cysteine biosynthesis in Arabidopsis.
Plant Cell Environ 2009; 32(4):349-67PC

Abstract

In plants, the enzymes for cysteine synthesis serine acetyltransferase (SAT) and O-acetylserine-(thiol)-lyase (OASTL) are present in the cytosol, plastids and mitochondria. However, it is still not clearly resolved to what extent the different compartments are involved in cysteine biosynthesis and how compartmentation influences the regulation of this biosynthetic pathway. To address these questions, we analysed Arabidopsis thaliana T-DNA insertion mutants for cytosolic and plastidic SAT isoforms. In addition, the subcellular distribution of enzyme activities and metabolite concentrations implicated in cysteine and glutathione biosynthesis were revealed by non-aqueous fractionation (NAF). We demonstrate that cytosolic SERAT1.1 and plastidic SERAT2.1 do not contribute to cysteine biosynthesis to a major extent, but may function to overcome transport limitations of O-acetylserine (OAS) from mitochondria. Substantiated by predominantly cytosolic cysteine pools, considerable amounts of sulphide and presence of OAS in the cytosol, our results suggest that the cytosol is the principal site for cysteine biosynthesis. Subcellular metabolite analysis further indicated efficient transport of cysteine, gamma-glutamylcysteine and glutathione between the compartments. With respect to regulation of cysteine biosynthesis, estimation of subcellular OAS and sulphide concentrations established that OAS is limiting for cysteine biosynthesis and that SAT is mainly present bound in the cysteine-synthase complex.

Authors+Show Affiliations

Max Planck Institut für Molekulare Pflanzenphysiologie, Am Mühlenberg 1, 14476 Potsdam-Golm, France.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19143986

Citation

Krueger, Stephan, et al. "Analysis of Cytosolic and Plastidic Serine Acetyltransferase Mutants and Subcellular Metabolite Distributions Suggests Interplay of the Cellular Compartments for Cysteine Biosynthesis in Arabidopsis." Plant, Cell & Environment, vol. 32, no. 4, 2009, pp. 349-67.
Krueger S, Niehl A, Lopez Martin MC, et al. Analysis of cytosolic and plastidic serine acetyltransferase mutants and subcellular metabolite distributions suggests interplay of the cellular compartments for cysteine biosynthesis in Arabidopsis. Plant Cell Environ. 2009;32(4):349-67.
Krueger, S., Niehl, A., Lopez Martin, M. C., Steinhauser, D., Donath, A., Hildebrandt, T., ... Hesse, H. (2009). Analysis of cytosolic and plastidic serine acetyltransferase mutants and subcellular metabolite distributions suggests interplay of the cellular compartments for cysteine biosynthesis in Arabidopsis. Plant, Cell & Environment, 32(4), pp. 349-67. doi:10.1111/j.1365-3040.2008.01928.x.
Krueger S, et al. Analysis of Cytosolic and Plastidic Serine Acetyltransferase Mutants and Subcellular Metabolite Distributions Suggests Interplay of the Cellular Compartments for Cysteine Biosynthesis in Arabidopsis. Plant Cell Environ. 2009;32(4):349-67. PubMed PMID: 19143986.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Analysis of cytosolic and plastidic serine acetyltransferase mutants and subcellular metabolite distributions suggests interplay of the cellular compartments for cysteine biosynthesis in Arabidopsis. AU - Krueger,Stephan, AU - Niehl,Annette, AU - Lopez Martin,M Carmen, AU - Steinhauser,Dirk, AU - Donath,Andrea, AU - Hildebrandt,Tatjana, AU - Romero,Luis C, AU - Hoefgen,Rainer, AU - Gotor,Cecilia, AU - Hesse,Holger, PY - 2009/1/16/entrez PY - 2009/1/16/pubmed PY - 2009/3/27/medline SP - 349 EP - 67 JF - Plant, cell & environment JO - Plant Cell Environ. VL - 32 IS - 4 N2 - In plants, the enzymes for cysteine synthesis serine acetyltransferase (SAT) and O-acetylserine-(thiol)-lyase (OASTL) are present in the cytosol, plastids and mitochondria. However, it is still not clearly resolved to what extent the different compartments are involved in cysteine biosynthesis and how compartmentation influences the regulation of this biosynthetic pathway. To address these questions, we analysed Arabidopsis thaliana T-DNA insertion mutants for cytosolic and plastidic SAT isoforms. In addition, the subcellular distribution of enzyme activities and metabolite concentrations implicated in cysteine and glutathione biosynthesis were revealed by non-aqueous fractionation (NAF). We demonstrate that cytosolic SERAT1.1 and plastidic SERAT2.1 do not contribute to cysteine biosynthesis to a major extent, but may function to overcome transport limitations of O-acetylserine (OAS) from mitochondria. Substantiated by predominantly cytosolic cysteine pools, considerable amounts of sulphide and presence of OAS in the cytosol, our results suggest that the cytosol is the principal site for cysteine biosynthesis. Subcellular metabolite analysis further indicated efficient transport of cysteine, gamma-glutamylcysteine and glutathione between the compartments. With respect to regulation of cysteine biosynthesis, estimation of subcellular OAS and sulphide concentrations established that OAS is limiting for cysteine biosynthesis and that SAT is mainly present bound in the cysteine-synthase complex. SN - 1365-3040 UR - https://www.unboundmedicine.com/medline/citation/19143986/Analysis_of_cytosolic_and_plastidic_serine_acetyltransferase_mutants_and_subcellular_metabolite_distributions_suggests_interplay_of_the_cellular_compartments_for_cysteine_biosynthesis_in_Arabidopsis_ L2 - https://doi.org/10.1111/j.1365-3040.2008.01928.x DB - PRIME DP - Unbound Medicine ER -