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Published on August 27, 2008; 10.1104/pp.108.125237

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Received June 24, 2008
Accepted August 25, 2008

Mitochondrial serine acetyltransferase functions as pacemaker of cysteine synthesis in plant cells

Florian H. Haas , Corinna Heeg , Rafael Queiroz , Andrea Bauer , Markus Wirtz , and Rudiger Hell *

Heidelberg Institute for Plant Sciences (HIP), Heidelberg University, Im Neuenheimer Feld 360, 69120 Heidelberg, Germany; German Cancer Research Center (DKFZ), Im Neuenheimer Feld 580, 69120 Heidelberg, Germany

* Corresponding author; email: rhell{at}hip.uni-heidelberg.de.

Cysteine synthesis in plants is carried out by two sequential reactions catalyzed by the rate-limiting enzyme serine acetyltransferase (SAT) and excess amounts of O-acetylserine(thiol)lyase (OAS-TL). Why these reactions occur in plastids, mitochondria and cytosol of plants remained unclear. Expression of artificial micro RNA (amiRNA) against Sat3 encoding mitochondrial SAT3 in transgenic Arabidopsis thaliana plants demonstrates that mitochondria are the most important compartment for synthesis of O-acetylserine (OAS), the precursor of cysteine. Reduction of RNA levels, protein contents, SAT enzymatic activity and phenotype strongly correlate in independent amiSAT3 lines and cause significantly retarded growth. The expression of the other four Sat genes in the Arabidopsis genome are not affected by amiRNA-SAT3 according to quantitative real time PCR and microarray analyses. Application of radiolabeled serine to leaf pieces reveales severely reduced incorporation rates into cysteine and even more so into glutathione. Accordingly steady-state levels of OAS are 4-fold reduced. Decrease of sulfate reduction related genes is accompanied by accumulation of sulfate in amiSAT3 lines. The results unequivocally show that mitochondria provide the bulk of OAS in the plant cell and are the likely site of flux regulation. Together with recent data [Heeg et al. (2008) Plant Cell 20: 168] the cytosol appears as major site of cysteine synthesis while plastids contribute reduced sulfur as sulfide. Thus, cysteine synthesis in plants is significantly different from that in non-photosynthetic eukaryotes at the cellular level.




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