Identification of an Arabidopsis unknown small membrane protein targeted to mitochondria, chloroplasts, and peroxisomes

Abu-Abied, Mohamad; Avisar, Dror; Belausov, Eduard; Holdengreber, Vered; Kam, Zvi; Sadot, Einat

doi:10.1007/s00709-009-0038-7

Original Article
Published: 13 March 2009

Identification of an Arabidopsis unknown small membrane protein targeted to mitochondria, chloroplasts, and peroxisomes

Mohamad Abu-Abied¹,
Dror Avisar¹,
Eduard Belausov¹,
Vered Holdengreber¹,
Zvi Kam² &
Einat Sadot¹

Protoplasma volume 236, pages 3–12 (2009)Cite this article

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Abstract

In a functional genomic screen performed by combining an Arabidopsis–yellow fluorescent protein (YFP)-fused complementary DNA (cDNA) library, rat fibroblasts as host and automatic microscopy, we found a short protein with a predictable trans-membrane domain encoded on chromosome 2. In rat fibroblasts, its pattern of distribution was to various organelle-like structures. From the databases, we learned that it has another family member in Arabidopsis and homologs in several other plants, Chlamydomonas and fungi, with a highly conserved N-terminal region. We named this protein from Arabidopsis short membrane protein (SMP) 2. No SMP homologs were found in mammalian sequence databases. When the full-length cDNAs of SMP2 was fused to YFP under the 35S promoter, comparable distribution was observed in Nicotiana benthamiana leaves, suggesting an unknown, evolutionarily conserved localization signal. Similar localization was observed when SMP2 was expressed in N. benthamiana leaves under the control of its own 5′ regulatory sequences. Colocalization studies with green fluorescent protein and red fluorescent protein chimeras revealed its colocalization with chloroplasts, peroxisomes, and mitochondria. No localization of SMP2 was observed in the Golgi. Immunostaining with specific antibodies corroborated the SMP2 localization to the three organelles.

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Abbreviations

SMP:: Small membrane protein
GFP:: Green fluorescent protein
RFP:: Red fluorescent protein
YFP:: Yellow fluorescent protein

References

Abu-Abied M, Golomb L, Belausov E, Huang S, Geiger B, Kam Z, Staiger CJ, Sadot E (2006) Identification of plant cytoskeleton-interacting proteins by screening for actin stress fiber association in mammalian fibroblasts. Plant J 48:367–379. doi:10.1111/j.1365-313X.2006.02883.x
PubMed Article CAS Google Scholar
Amery L, Fransen M, De Nys K, Mannaerts GP, Van Veldhoven PP (2000) Mitochondrial and peroxisomal targeting of 2-methylacyl-CoA racemase in humans. J Lipid Res 41:1752–1759
PubMed CAS Google Scholar
Baker A, Sparkes IA (2005) Peroxisome protein import: some answers, more questions. Curr Opin Plant Biol 8:640–647. doi:10.1016/j.pbi.2005.09.009
PubMed Article CAS Google Scholar
Brugiere S, Kowalski S, Ferro M, Seigneurin-Berny D, Miras S, Salvi D, Ravanel S, d'Herin P, Garin J, Bourguignon J, Joyard J, Rolland N (2004) The hydrophobic proteome of mitochondrial membranes from Arabidopsis cell suspensions. Phytochemistry 65:1693–1707. doi:10.1016/j.phytochem.2004.03.028
PubMed Article CAS Google Scholar
Chew O, Whelan J, Millar AH (2003) Molecular definition of the ascorbate–glutathione cycle in Arabidopsis mitochondria reveals dual targeting of antioxidant defenses in plants. J Biol Chem 278:46869–46877. doi:10.1074/jbc.M307525200
PubMed Article CAS Google Scholar
Chew O, Lister R, Qbadou S, Heazlewood JL, Soll J, Schleiff E, Millar AH, Whelan J (2004) A plant outer mitochondrial membrane protein with high amino acid sequence identity to a chloroplast protein import receptor. FEBS Lett 557:109–114. doi:10.1016/S0014-5793(03)01457-1
PubMed Article CAS Google Scholar
Christensen AC, Lyznik A, Mohammed S, Elowsky CG, Elo A, Yule R, Mackenzie SA (2005) Dual-domain, dual-targeting organellar protein presequences in Arabidopsis can use non-AUG start codons. Plant Cell 17:2805–2816. doi:10.1105/tpc.105.035287
PubMed Article CAS Google Scholar
Dhonukshe P, Samaj J, Baluska F, Friml J (2007) A unifying new model of cytokinesis for the dividing plant and animal cells. Bioessays 29:371–381. doi:10.1002/bies.20559
PubMed Article CAS Google Scholar
Duchene AM, Giritch A, Hoffmann B, Cognat V, Lancelin D, Peeters NM, Zaepfel M, Marechal-Drouard L, Small ID (2005) Dual targeting is the rule for organellar aminoacyl-tRNA synthetases in Arabidopsis thaliana. Proc Natl Acad Sci U S A 102:16484–16489. doi:10.1073/pnas.0504682102
PubMed Article CAS Google Scholar
Elo A, Lyznik A, Gonzalez DO, Kachman SD, Mackenzie SA (2003) Nuclear genes that encode mitochondrial proteins for DNA and RNA metabolism are clustered in the Arabidopsis genome. Plant Cell 15:1619–1631. doi:10.1105/tpc.010009
PubMed Article CAS Google Scholar
Heazlewood JL, Verboom RE, Tonti-Filippini J, Small I, Millar AH (2007) SUBA: the Arabidopsis subcellular database. Nucleic Acids Res 35:D213–D218. doi:10.1093/nar/gkl863
PubMed Article CAS Google Scholar
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685. doi:10.1038/227680a0
PubMed Article CAS Google Scholar
Lister R, Carrie C, Duncan O, Ho LH, Howell KA, Murcha MW, Whelan J (2007) Functional definition of outer membrane proteins involved in preprotein import into mitochondria. Plant Cell 19:3739–3759. doi:10.1105/tpc.107.050534
PubMed Article CAS Google Scholar
Logan DC, Leaver CJ (2000) Mitochondria-targeted GFP highlights the heterogeneity of mitochondrial shape, size and movement within living plant cells. J Exp Bot 51:865–871. doi:10.1093/jexbot/51.346.865
PubMed Article CAS Google Scholar
Lunn JE (2007) Compartmentation in plant metabolism. J Exp Bot 58:35–47. doi:10.1093/jxb/erl134
PubMed Article CAS Google Scholar
Mackenzie SA (2005) Plant organellar protein targeting: a traffic plan still under construction. Trends Cell Biol 15:548–554. doi:10.1016/j.tcb.2005.08.007
PubMed Article CAS Google Scholar
Meyerowitz EM (2002) Plants compared to animals: the broadest comparative study of development. Science 295:1482–1485. doi:10.1126/science.1066609
PubMed Article CAS Google Scholar
Millar AH, Whelan J, Small I (2006) Recent surprises in protein targeting to mitochondria and plastids. Curr Opin Plant Biol 9:610–615. doi:10.1016/j.pbi.2006.09.002
PubMed Article CAS Google Scholar
Nelson BK, Cai X, Nebenfuhr A (2007) A multicolored set of in vivo organelle markers for co-localization studies in Arabidopsis and other plants. Plant J 51:1126–1136. doi:10.1111/j.1365-313X.2007.03212.x
PubMed Article CAS Google Scholar
Oreb M, Tews I, Schleiff E (2008) Policing Tic 'n' Toc, the doorway to chloroplasts. Trends Cell Biol 18:19–27. doi:10.1016/j.tcb.2007.10.002
PubMed Article CAS Google Scholar
Peeters N, Small I (2001) Dual targeting to mitochondria and chloroplasts. Biochim Biophys Acta 1541:54–63. doi:10.1016/S0167-4889(01)00146-X
PubMed Article CAS Google Scholar
Perry AJ, Rimmer KA, Mertens HD, Waller RF, Mulhern TD, Lithgow T, Gooley PR (2008) Structure, topology and function of the translocase of the outer membrane of mitochondria. Plant Physiol Biochem 46:265–274. doi:10.1016/j.plaphy.2007.12.012
PubMed Article CAS Google Scholar
Petrova VY, Drescher D, Kujumdzieva AV, Schmitt MJ (2004) Dual targeting of yeast catalase A to peroxisomes and mitochondria. Biochem J 380:393–400. doi:10.1042/BJ20040042
PubMed Article CAS Google Scholar
Rassow J, Dekker PJ, van Wilpe S, Meijer M, Soll J (1999) The preprotein translocase of the mitochondrial inner membrane: function and evolution. J Mol Biol 286:105–120. doi:10.1006/jmbi.1998.2455
PubMed Article CAS Google Scholar
Sablowski R (2004) Plant and animal stem cells: conceptually similar, molecularly distinct? Trends Cell Biol 14:605–611. doi:10.1016/j.tcb.2004.09.011
PubMed Article CAS Google Scholar
Saint-Jore-Dupas C, Nebenfuhr A, Boulaflous A, Follet-Gueye ML, Plasson C, Hawes C, Driouich A, Faye L, Gomord V (2006) Plant N-glycan processing enzymes employ different targeting mechanisms for their spatial arrangement along the secretory pathway. Plant Cell 18:3182–3200. doi:10.1105/tpc.105.036400
PubMed Article CAS Google Scholar
Sanderson MJ (2008) Phylogenetic signal in the eukaryotic tree of life. Science 321:121–123. doi:10.1126/science.1154449
PubMed Article CAS Google Scholar
Shaner NC, Campbell RE, Steinbach PA, Giepmans BN, Palmer AE, Tsien RY (2004) Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nat Biotechnol 22:1567–1572. doi:10.1038/nbt1037
PubMed Article CAS Google Scholar
Silva-Filho MC (2003) One ticket for multiple destinations: dual targeting of proteins to distinct subcellular locations. Curr Opin Plant Biol 6:589–595. doi:10.1016/j.pbi.2003.09.008
PubMed Article CAS Google Scholar
Sunderhaus S, Dudkina NV, Jansch L, Klodmann J, Heinemeyer J, Perales M, Zabaleta E, Boekema EJ, Braun HP (2006) Carbonic anhydrase subunits form a matrix-exposed domain attached to the membrane arm of mitochondrial complex I in plants. J Biol Chem 281:6482–6488. doi:10.1074/jbc.M511542200
PubMed Article CAS Google Scholar
Voigt B, Timmers AC, Samaj J, Hlavacka A, Ueda T, Preuss M, Nielsen E, Mathur J, Emans N, Stenmark H, Nakano A, Baluska F, Menzel D (2005) Actin-based motility of endosomes is linked to the polar tip growth of root hairs. Eur J Cell Biol 84:609–621. doi:10.1016/j.ejcb.2004.12.029
PubMed Article CAS Google Scholar
Zimmermann P, Hirsch-Hoffmann M, Hennig L, Gruissem W (2004) GENEVESTIGATOR. Arabidopsis microarray database and analysis toolbox. Plant Physiol 136:2621–2632. doi:10.1104/pp. 104.046367
PubMed Article CAS Google Scholar

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Acknowledgments

The authors thank Yael Paran, Irena Lavelin, and Yuvalal Lyron for their help with the automatic microscope and Dr. Oren Ostersetzer-Biran for critical reading of the manuscript. This research was supported by a research grant from the Israeli Science Foundation (ISF) #752/05.

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Affiliations

The Department of Ornamental Horticulture, The Institute of Plant Sciences, The Volcani Center, P.O. Box 6, Bet-Dagan, 50250, Israel
Mohamad Abu-Abied, Dror Avisar, Eduard Belausov, Vered Holdengreber & Einat Sadot
The Department of Molecular Biology of the Cell, The Weizmann Institute of Science, Rehovot, 76100, Israel
Zvi Kam

Authors

Mohamad Abu-Abied
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Dror Avisar
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Eduard Belausov
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Vered Holdengreber
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Zvi Kam
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Einat Sadot
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Corresponding author

Correspondence to Einat Sadot.

Electronic Supplementary Material

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Supplementary Fig. 1

Expression profiles of SMP1 and SMP2 according to the Genevestigator (XLS 26 kb).

High resolution image file (TIFF 5.82 MB).

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Abu-Abied, M., Avisar, D., Belausov, E. et al. Identification of an Arabidopsis unknown small membrane protein targeted to mitochondria, chloroplasts, and peroxisomes. Protoplasma 236, 3–12 (2009). https://doi.org/10.1007/s00709-009-0038-7

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Received: 15 February 2009
Accepted: 23 February 2009
Published: 13 March 2009
Issue Date: July 2009
DOI: https://doi.org/10.1007/s00709-009-0038-7

Keywords

Arabidopsis
Chloroplasts
Mitochondria
Peroxisomes
GFP