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A plastidial sodium-dependent pyruvate transporter

A Corrigendum to this article was published on 28 September 2011

This article has been updated

Abstract

Pyruvate serves as a metabolic precursor for many plastid-localized biosynthetic pathways, such as those for fatty acids1, terpenoids2 and branched-chain amino acids3. In spite of the importance of pyruvate uptake into plastids (organelles within cells of plants and algae), the molecular mechanisms of this uptake have not yet been explored. This is mainly because pyruvate is a relatively small compound that is able to passively permeate lipid bilayers4, which precludes accurate measurement of pyruvate transport activity in reconstituted liposomes. Using differential transcriptome analyses of C3 and C4 plants of the genera Flaveria and Cleome, here we have identified a novel gene that is abundant in C4 species, named BASS2 (BILE ACID:SODIUM SYMPORTER FAMILY PROTEIN 2). The BASS2 protein is localized at the chloroplast envelope membrane, and is highly abundant in C4 plants that have the sodium-dependent pyruvate transporter. Recombinant BASS2 shows sodium-dependent pyruvate uptake activity. Sodium influx is balanced by a sodium:proton antiporter (NHD1), which was mimicked in recombinant Escherichia coli cells expressing both BASS2 and NHD1. Arabidopsis thaliana bass2 mutants lack pyruvate uptake into chloroplasts, which affects plastid-localized isopentenyl diphosphate synthesis, as evidenced by increased sensitivity of such mutants to mevastatin, an inhibitor of cytosolic isopentenyl diphosphate biosynthesis. We thus provide molecular evidence for a sodium-coupled metabolite transporter in plastid envelopes. Orthologues of BASS2 can be detected in all the genomes of land plants that have been characterized so far, thus indicating the widespread importance of sodium-coupled pyruvate import into plastids.

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Figure 1: The novel C 4 -associated transcripts encoding plastid-targeted membrane proteins.
Figure 2: BASS2 protein levels.
Figure 3: Pyruvate uptake activity.
Figure 4: BASS2 function in A. thaliana.

Accession codes

Primary accessions

GenBank/EMBL/DDBJ

Data deposits

cDNA sequences for F. trinervia BASS2, F. trinervia BASS4 and F. bidentis NHD1 have been deposited in the DNA Data Bank of Japan, with respective accession numbers AB522102, AB522103 and AB642169.

Change history

  • 17 October 2011

    Parts of the Supplementary Information were inadvertently not uploaded and the file was also corrupted; this has been corrected.

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Acknowledgements

We thank G. Schönknecht, S. Yamaguchi and Y. Kamiya for discussions; N. Das, S. von Caemmerer and R. T. Furbank for critical reading of the manuscript; R. F. Sage, T. Endo, M. Munekage, J. Hibberd and M. Ku for gifts of seeds; T. Kinoshita for technical advice on the BASS2 immunoblot analysis; N. Aoki and S. Koreeda for technical advice on the pyruvate-uptake measurements; A. Izumida for preparation of the F. trinervia cDNA library; and Y. Takahashi for suggesting the dual expression system. This work was supported in part by the Ministry of Education, Science and Culture of Japan (Grants-in-Aid for Scientific Research to T.F. and K.I.), by a Sasakawa Scientific Research Grant from the Japan Science Society to T.F., and by the German Research Foundation (CRC-TR1 and IRTG 1525/1 to A.P.M.W., and CRC 590 to P.W.).

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Authors

Contributions

T.Y. performed the differential screening and isolated the BASS2 gene. M.N. and Y.O.-I. analysed C4-abundant genes. Y.O.-I. prepared the bass2 mutants and revealed BASS2 expression. M.S. performed the confocal laser micro-scanning and the analyses of immunohistochemistry. J.O. and T.F. performed the pyruvate-uptake measurements on isolated chloroplasts, and A.B., A.P.M.W. and T.F. performed the E. coli whole-cell uptake measurements. U.G., P.W., A.B. and A.P.M.W. performed Flaveria and Cleome transcriptome analyses and the phylogenic analysis. S.H. was involved in designing the study, T.F. and K.I. designed the study, and T.F. and A.P.M.W. wrote the paper.

Corresponding author

Correspondence to Tsuyoshi Furumoto.

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The authors declare no competing financial interests.

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The file contains Supplementary Text, Supplementary References and Supplementary Figures 1-12 with legends. This file was replaced on 17 October 2011 as the Supplementary Text and Supplementary References were omitted from the original file posted on line. (PDF 4233 kb)

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Furumoto, T., Yamaguchi, T., Ohshima-Ichie, Y. et al. A plastidial sodium-dependent pyruvate transporter. Nature 476, 472–475 (2011). https://doi.org/10.1038/nature10250

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