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Plant Molecular Biology

, Volume 88, Issue 1–2, pp 3–20 | Cite as

Multiple internal sorting determinants can contribute to the trafficking of cruciferin to protein storage vacuoles

  • Dwayne D. Hegedus
  • Cathy Coutu
  • Myrtle Harrington
  • Brad Hope
  • Kelsey Gerbrandt
  • Ivo Nikolov
Article

Abstract

Trafficking of seed storage proteins to protein storage vacuoles is mediated by carboxy terminal and internal sorting determinants (ISDs). Protein modelling was used to identify candidate ISDs residing near surface-exposed regions in Arabidopsis thaliana cruciferin A (AtCruA). These were verified by AtCruA fusion to yellow fluorescent protein (YFP) and expression in developing embryos of A. thaliana. As the presence of endogenous cruciferin was found to mask the effects of weaker ISDs, experiments were conducted in a line that was devoid of cruciferin. In total, nine ISDs were discovered and a core determinant defined using a series of alanine scanning and deletion mutant variants. Coupling of functional data from AtCruA ISD-YFP fusions with statistical analysis of the physiochemical properties of analogous regions from several 11/12S globulins revealed that cruciferin ISDs likely adhere to the following rules: (1) ISDs are adjacent to or within hydrophilic, surface-exposed regions that serve to present them on the protein’s surface; (2) ISDs generally have a hydrophobic character; (3) ISDs tend to have Leu or Ile residues at their core; (4) ISDs are approximately eight amino acids long with the physiochemical consensus [hydrophobic][preferably charged][small or hydrophobic, but not tiny][IL][polar, preferably charged][small, but not charged][hydrophobic, not charged, preferably not polar][hydrophobic, not tiny, preferably not polar]. Microscopic evidence is also presented for the presence of an interconnected protein storage vacuolar network in embryo cells, rather than discreet, individual vacuoles.

Keywords

Seed storage protein Cruciferin Sorting determinants Protein storage vacuole 

Notes

Acknowledgments

This work was supported with funding from the Agriculture and Agri-Food Canada Canadian Crop Genomics Initiative.

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References

  1. Adachi M, Takenaka Y, Gidamis AB, Mikami B, Utsumi S (2001) Crystal structure of soybean proglycinin A1aB1b homotrimer. J Mol Biol 305:291–305CrossRefPubMedGoogle Scholar
  2. Adachi M, Kanamori J, Masuda T, Yagasaki K, Kitamura K, Mikami B, Utsumi S (2003) Crystal structure of soybean 11S globulin: glycinin A3B4 homohexamer. Proc Natl Acad Sci USA 100:7395–7400CrossRefPubMedCentralPubMedGoogle Scholar
  3. Arnold K, Bordoli L, Kopp J, Schwede T (2006) The SWISS-MODEL workspace: a web-based environment for protein structure homology modelling. Bioinformatics 22:195–201CrossRefPubMedGoogle Scholar
  4. Bechtold N, Ellis J, Pelletier F (1993) In planta Agrobacterium mediated gene transfer by infiltration of adult Arabidopsis thaliana plants. C R Acad Sci Paris Life Sci 316:1194–1199Google Scholar
  5. Brown JC, Jolliffe NA, Frigerio L, Roberts LM (2003) Sequence-specific, Golgi-dependent vacuolar targeting of castor bean 2S albumin. Plant J 36:711–719CrossRefPubMedGoogle Scholar
  6. Chrispeels MJ (1983) The Golgi apparatus mediates the transport of phytoheagglutinin to the protein bodies in bean cotyledons. Planta 158:140–151CrossRefPubMedGoogle Scholar
  7. Chrispeels MJ, Higgins TJ, Spencer D (1982) Assembly of storage protein oligomers in the endoplasmic reticulum and processing of the polypeptides in the protein bodies of developing pea cotyledons. J Cell Biol 93:306–313CrossRefPubMedCentralPubMedGoogle Scholar
  8. Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743CrossRefPubMedGoogle Scholar
  9. Craddock CP, Hunter PR, Szakacs E, Hinz G, Robinson DG, Frigerio L (2008) Lack of a vacuolar sorting receptor leads to non-specific missorting of soluble vacuolar proteins in Arabidopsis seeds. Traffic 9:408–416CrossRefPubMedGoogle Scholar
  10. Craig S (1986) Fixation of a vacuole-associated network of channels in protein-storing pea cotyledon cells. Protoplasma 135:67–70CrossRefGoogle Scholar
  11. Curtis MD, Grossniklaus U (2003) A Gateway cloning vector set for high-throughput functional analysis of genes in planta. Plant Physiol 133:462–469CrossRefPubMedCentralPubMedGoogle Scholar
  12. Ebine K, Okatani Y, Uemura T, Goh T, Shoda K, Niihama M, Morita MT, Spitzer C, Otegui MS, Nakano A, Ueda T (2008) A SNARE complex unique to seed plants is required for protein storage vacuole biogenesis and seed development of Arabidopsis thaliana. Plant Cell 20:3006–3021CrossRefPubMedCentralPubMedGoogle Scholar
  13. Eddy SR (1998) Profile hidden Markov models. Bioinformatics 14:755–763CrossRefPubMedGoogle Scholar
  14. Frigerio L, de Virgilio M, Prada A, Faoro F, Vitale A (1998) Sorting of phaseolin to the vacuole is saturable and requires a short C-terminal peptide. Plant Cell 10:1031–1042CrossRefPubMedCentralPubMedGoogle Scholar
  15. Frigerio L, Jolliffe NA, Di Cola A, Felipe DH, Paris N, Neuhaus JM, Lord JM, Ceriotti A, Roberts LM (2001) The internal propeptide of the ricin precursor carries a sequence-specific determinant for vacuolar sorting. Plant Physiol 126:167–175CrossRefPubMedCentralPubMedGoogle Scholar
  16. Frigerio L, Hinz G, Robinson DG (2008) Multiple vacuoles in plant cells: rule or exception. Traffic 9:1564–1570CrossRefPubMedGoogle Scholar
  17. Hanton SL, Brandizzi F (2006) Post-Golgi protein traffic in the plant secretory pathway. Microsc Res Tech 69:152–159CrossRefPubMedGoogle Scholar
  18. Hanton SL, Matheson LA, Chatre L, Rossi M, Brandizzi F (2007) Post-Golgi protein traffic in the plant secretory pathway. Plant Cell Rep 26:1431–1438CrossRefPubMedGoogle Scholar
  19. Hara-Nishimura I, Shimada T, Hatano K, Takeuchi Y, Nishimura M (1998) Transport of storage proteins to protein storage vacuoles is mediated by large precursor-accumulating vesicles. Plant Cell 10:825–836CrossRefPubMedCentralPubMedGoogle Scholar
  20. Hayashi M, Toriyama K, Kondo M, Hara-Nishimura I, Nishimura M (1999) Accumulation of a fusion protein containing 2S albumin induces novel vesicles in vegetative cells of Arabidopsis. Plant Cell Physiol 40:263–272CrossRefPubMedGoogle Scholar
  21. Herman E, Schmidt M (2004) Endoplasmic reticulum to vacuole trafficking of endoplasmic reticulum bodies provides an alternate pathway for protein transfer to vacuoles. Plant Physiol 136:3440–3446CrossRefPubMedCentralPubMedGoogle Scholar
  22. Hinz G, Hillmer S, Bäumer M, Hohl I (1999) Vacuolar storage proteins and the putative vacuolar sorting receptor BP-80 exit the Golgi apparatus of developing pea cotyledons in different transport vesicles. Plant Cell 11:1509–1524CrossRefPubMedCentralPubMedGoogle Scholar
  23. Hinz G, Colanesi S, Hillmer S, Rogers JC, Robinson DG (2007) Localization of vacuolar transport receptors and cargo proteins in the Golgi apparatus of developing Arabidopsis embryos. Traffic 8:1452–1464CrossRefPubMedGoogle Scholar
  24. Hoh B, Hinz G, Jeong BK, Robinson DG (1995) Protein storage vacuoles form de novo during pea cotyledon development. J Cell Sci 108:299–310PubMedGoogle Scholar
  25. Hohl I, Robinson DG, Chrispeels MJ, Hinz G (1996) Transport of storage proteins to the vacuole is mediated by vesicles without a clathrin coat. J Cell Sci 109:2539–2550PubMedGoogle Scholar
  26. Holkeri H, Vitale A (2001) Vacuolar sorting determinants within a plant storage protein trimer act cooperatively. Traffic 2:737–741CrossRefPubMedGoogle Scholar
  27. Horton RM, Cai ZL, Ho SN, Pease LR (1990) Gene splicing by overlap extension: Tailor-made genes using the polymerase chain reaction. Biotechniques 8:528–535PubMedGoogle Scholar
  28. Hunter PR, Craddock CP, Di Benedetto S, Roberts LM, Frigerio L (2007) Fluorescent reporter proteins for the tonoplast and the vacuolar lumen identify a single vacuolar compartment in Arabidopsis cells. Plant Physiol 145:1371–1382CrossRefPubMedCentralPubMedGoogle Scholar
  29. Inoue S (2006) Foundations of confocal scanned imaging in light microscopy. In: JB Pawley (ed) Handbook of biological confocal microscopy, 3rd Edn.   Springer Science and Business Media, LLC.Google Scholar
  30. Jiang L, Phillips TE, Rogers SW, Rogers JC (2000) Biogenesis of the protein storage vacuole crystalloid. J Cell Biol 150:755–769CrossRefPubMedCentralPubMedGoogle Scholar
  31. Jiang L, Phillips TE, Hamm CA, Drozdowicz YM, Rea PA, Maeshima M, Rogers SW, Rogers JC (2001) The protein storage vacuole: a unique compound organelle. J Cell Biol 155:991–1002CrossRefPubMedCentralPubMedGoogle Scholar
  32. Jolliffe NA, Brown JC, Neumann U, Vicré M, Bachi A, Hawes C, Ceriotti A, Roberts LM, Frigerio L (2004) Transport of ricin and 2S albumin precursors to the storage vacuoles of Ricinus communis endosperm involves the Golgi and VSR-like receptors. Plant J 39:821–833CrossRefPubMedGoogle Scholar
  33. Jolliffe NA, Craddock CP, Frigerio L (2005) Pathways for protein transport to seed storage vacuoles. Biochem Soc Trans 33:1016–1018CrossRefPubMedGoogle Scholar
  34. Jung R, Scott MP, Nam YW, Beaman TW, Bassüner R, Saalbach I, Müntz K, Nielsen NC (1998) The role of proteolysis in the processing and assembly of 11S seed globulins. Plant Cell 10:343–357CrossRefPubMedCentralPubMedGoogle Scholar
  35. Livingstone C, Barton G (1993) Protein sequence alignments: a strategy for hierarchical analysis of residue conservation. Comp Appl Biosci 9:745–756PubMedGoogle Scholar
  36. Maruyama N, Mun LC, Tatsuhara M, Sawada M, Ishimoto M, Utsumi S (2006) Multiple vacuolar sorting determinants exist in soybean 11S globulin. Plant Cell 18:1253–1273CrossRefPubMedCentralPubMedGoogle Scholar
  37. Matsuoka K, Nakamura K (1999) Large alkyl side-chains of isoleucine and leucine in the NPIRL region constitute the core of the vacuolar sorting determinant of sporamin precursor. Plant Mol Biol 41:825–835CrossRefPubMedGoogle Scholar
  38. Matsuoka K, Neuhaus J-M (1999) Cis-elements of protein transport to the plant vacuoles. J Exp Biol 50:165–174Google Scholar
  39. Mori T, Maruyama N, Nishizawa K, Higasa T, Yagasaki K, Ishimoto M, Utsumi S (2004) The composition of newly synthesized proteins in the endoplasmic reticulum determines the transport pathways of soybean seed storage proteins. Plant J 40:238–249CrossRefPubMedGoogle Scholar
  40. Nishizawa K, Maruyama N, Satoh R, Fuchikami Y, Higasa T, Utsumi S (2003) A C-terminal sequence of soybean beta-conglycinin alpha’ subunit acts as a vacuolar sorting determinant in seed cells. Plant J 34:647–659CrossRefPubMedGoogle Scholar
  41. Nishizawa K, Maruyama N, Satoh R, Higasa T, Utsumi S (2004) A vacuolar sorting determinant of soybean β-conglycinin β subunit resides in a C-terminal sequence. Plant Sci 167:937–947CrossRefGoogle Scholar
  42. Nishizawa K, Maruyama N, Utsumi S (2006) The C-terminal region of alpha’ subunit of soybean beta-conglycinin contains two types of vacuolar sorting determinants. Plant Mol Biol 62:111–125CrossRefPubMedGoogle Scholar
  43. Otegui MS, Herder R, Schulze J, Jung R, Staehelin LA (2006) The proteolytic processing of seed storage proteins in Arabidopsis embryo cells starts in the multivesicular bodies. Plant Cell 18:2567–2581CrossRefPubMedCentralPubMedGoogle Scholar
  44. Paris N, Neuhaus J-M (2002) BP-80 as a vacuolar sorting receptor. Plant Mol Biol 50:903–914CrossRefPubMedGoogle Scholar
  45. Park M, Lee D, Lee G-J, Hwang I (2005) AtRMR1 functions as a cargo receptor for protein trafficking to the protein storage vacuole. J Cell Biol 170:757–767CrossRefPubMedCentralPubMedGoogle Scholar
  46. Park JH, Oufattole M, Rogers JC (2007) Golgi-mediated vacuolar sorting in plant cells: RMR proteins are sorting receptors for the protein aggregation/membrane internalization pathway. Plant Sci 172:728–745CrossRefGoogle Scholar
  47. Petruccelli S, Molina MI, Lareu FJ, Circosta A (2007) Two short sequences from amaranth 11S globulin are sufficient to target green fluorescent protein and beta-glucuronidase to vacuoles in Arabidopsis cells. Plant Physiol Biochem 45:400–409CrossRefPubMedGoogle Scholar
  48. Robinson DG, Bäumer M, Hinz G, Hohl I (1998) Vesicle transfer of storage proteins to the vacuole: the role of the Golgi apparatus and multivesicular bodies 152:659–667Google Scholar
  49. Robinson DG, Oliviusson P, Hinz G (2005) Protein sorting to the storage vacuoles of plants: a critical appraisal. Traffic 6:615–625CrossRefPubMedGoogle Scholar
  50. Shen Y, Wang J, Ding Y, Lo SW, Gouzerh G, Neuhasu J-M, Jiang L (2011) The rice RMR1 associates with a distinct prevacuolar compartment for the protein storage vacuole pathway. Mol Plant 4:854–868CrossRefPubMedGoogle Scholar
  51. Shewry PR, Napier JA, Tatham AS (1995) Seed storage proteins: structures and biosynthesis. Plant Cell 7:945–956CrossRefPubMedCentralPubMedGoogle Scholar
  52. Shimada T, Kuroyanagi M, Nishimura M, Hara-Nishimura I (1997) A pumpkin 72-kDa membrane protein of precursor-accumulating vesicles has characteristics of a vacuolar sorting receptor. Plant Cell Physiol 38:1414–1420CrossRefPubMedGoogle Scholar
  53. Shimada T, Watanabe E, Tamura K, Hayashi Y, Nishimura M, Hara-Nishimura I (2002) A vacuolar sorting receptor PV72 on the membrane of vesicles that accumulate precursors of seed storage proteins (PAC vesicles). Plant Cell Physiol 43:1086–1095CrossRefPubMedGoogle Scholar
  54. Shimada T, Fuji K, Tamura K, Kondo M, Nishimura M, Hara-Nishimura I (2003a) Vacuolar sorting receptor for seed storage proteins in Arabidopsis thaliana. Proc Natl Acad Sci USA 100:16095–16100CrossRefPubMedCentralPubMedGoogle Scholar
  55. Shimada T, Yamada K, Kataoka M, Nakaune S, Koumoto Y, Kuroyanagi M, Tabata S, Kato T, Shinozaki K, Seki M, Kobayashi M, Kondo M, Nishimura M, Hara-Nishimura I (2003b) Vacuolar processing enzymes are essential for proper processing of seed storage proteins in Arabidopsis thaliana. J Biol Chem 278:32292–32299CrossRefPubMedGoogle Scholar
  56. Tamura K, Takahashi H, Kunieda T, Fuji K, Shimada T, Hara-Nishimura I (2007) Arabidopsis KAM2/GRV2 is required for proper endosome formation and functions in vacuolar sorting and determination of the embryo growth axis. Plant Cell 19:320–332CrossRefPubMedCentralPubMedGoogle Scholar
  57. Tandang-Silvas MR, Fukuda T, Fukuda C, Prak K, Cabanos C, Kimura A, Itoh T, Mikami B, Utsumi S, Maruyama N (2010) Conservation and divergence on plant seed 11S globulins based on crystal structures. Biochim Biophys Acta 1804:1432–1442CrossRefPubMedGoogle Scholar
  58. Vitale A, Hinz G (2005) Sorting of proteins to storage vacuoles: how many mechanisms? Trends Plant Sci 10:316–323CrossRefPubMedGoogle Scholar
  59. Wan L, Ross ARS, Yang J, Hegedus DD, Kermode AR (2007) Phosphorylation of 12S globulin cruciferin in Arabidopsis thaliana seeds of wild type and abi1-1 mutant. Biochem J 404:247–256CrossRefPubMedCentralPubMedGoogle Scholar
  60. Wang J, Tse YC, Hinz G, Robinson DG, Jiang L (2012) Storage globulins pass through the Golgi apparatus and multivesicular bodies in the absence of dense vesicle formation during early stages of cotyledon development in mung bean. J Exp Bot 63:1367–1380CrossRefPubMedCentralPubMedGoogle Scholar
  61. Withana-Gamage TS, Hegedus DD, Qiu X, Wanasundara J (2011) In silico homology modeling to predict protein functional properties of cruciferin. J Agric Food Chem 59:12925–12938CrossRefPubMedGoogle Scholar
  62. Withana-Gamage TS, Hegedus DD, Qui X, Yu P, May T, Lydiate D, Wanasundara JPD (2013) Characterization of Arabidopsis thaliana lines with altered seed storage protein profiles using synchrotron powered FTIR. J Agric Food Chem 61:901–912CrossRefPubMedGoogle Scholar
  63. Zdobnov EM, Apweiler R (2001) InterProScan—an integration platform for the signature-recognition methods in InterPro. Bioinformatics 17:847–848CrossRefPubMedGoogle Scholar
  64. Zimmermann P, Hirsch-Hoffmann M, Hennig L, Gruissem W (2004) GENEVESTIGATOR. Arabidopsis microarray database and analysis toolbox. Plant Physiol 136:2621–2632CrossRefPubMedCentralPubMedGoogle Scholar
  65. Zouhar J, Rojo E (2009) Plant vacuoles: where did they come from and where are they heading? Curr Opin Plant Biol 12:677–684CrossRefPubMedGoogle Scholar
  66. Zouhar J, Munoz A, Rojo E (2010) Functional specialization within the vacuolar sorting receptor family: VSR1, VSR3 and VSR4 sort vacuolar storage cargo in seeds and vegetative tissues. Plant J 64:577–588CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Dwayne D. Hegedus
    • 1
    • 2
  • Cathy Coutu
    • 1
  • Myrtle Harrington
    • 1
  • Brad Hope
    • 1
    • 2
  • Kelsey Gerbrandt
    • 1
  • Ivo Nikolov
    • 1
  1. 1.Agriculture and Agri-Food CanadaSaskatoonCanada
  2. 2.Department of Food and Bioproduct SciencesUniversity of SaskatchewanSaskatoonCanada

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