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Plant Cell. 1997 Feb; 9(2): 185–197.
PMCID: PMC156910

Oilseed isocitrate lyases lacking their essential type 1 peroxisomal targeting signal are piggybacked to glyoxysomes.


Isocitrate lyase (IL) is an essential enzyme in the glyoxylate cycle, which is a pathway involved in the mobilization of stored lipids during postgerminative growth of oil-rich seedlings. We determined experimentally the necessary and sufficient peroxisome targeting signals (PTSs) for cottonseed, oilseed rape, and castor bean ILs in a well-characterized in vivo import system, namely, suspension-cultured tobacco (Bright Yellow) BY-2 cells. Results were obtained by comparing immunofluorescence localizations of wild-type and C-terminal-truncated proteins transiently expressed from cDNAs introduced by microprojectile bombardment. The tripeptides ARM-COOH (on cottonseed and castor bean ILs) and SRM-COOH (on oilseed rape IL) were necessary for targeting and actual import of these ILs into glyoxysomes, and ARM-COOH was sufficient for redirecting chloramphenicol acetyltransferase (CAT) from the cytosol into the glyoxysomes. Surprisingly, IL and CAT subunits without these tripeptides were still acquired by glyoxysomes, but only when wild-type IL or CAT-SKL subunits, respectively, were simultaneously expressed in the cells. These results reveal that targeting signal-depleted subunits are being piggybacked as multimers to glyoxysomes by association with subunits possessing a PTS1. Targeted multimers are then translocated through membrane pores or channels to the matrix as oligomers or as subunits before reoligomerization in the matrix.

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  • BANGHAM AD, HORNE RW, GLAUERT AM, DINGLE JT, LUCY JA. Action of saponin on biological cell membranes. Nature. 1962 Dec 8;196:952–955. [PubMed]
  • Banjoko A, Trelease RN. Development and application of an in vivo plant peroxisome import system. Plant Physiol. 1995 Apr;107(4):1201–1208. [PMC free article] [PubMed]
  • Behari R, Baker A. The carboxyl terminus of isocitrate lyase is not essential for import into glyoxysomes in an in vitro system. J Biol Chem. 1993 Apr 5;268(10):7315–7322. [PubMed]
  • Comai L, Dietrich RA, Maslyar DJ, Baden CS, Harada JJ. Coordinate expression of transcriptionally regulated isocitrate lyase and malate synthase genes in Brassica napus L. Plant Cell. 1989 Mar;1(3):293–300. [PMC free article] [PubMed]
  • Davis WL, Jones RG, Farmer GR, Matthews JL, Goodman DB. Glyoxylate cycle in the epiphyseal growth plate: isocitrate lyase and malate synthase identified in mammalian cartilage. Anat Rec. 1989 Apr;223(4):357–362. [PubMed]
  • Davis WL, Goodman DB, Crawford LA, Cooper OJ, Matthews JL. Hibernation activates glyoxylate cycle and gluconeogenesis in black bear brown adipose tissue. Biochim Biophys Acta. 1990 Mar 9;1051(3):276–278. [PubMed]
  • de Hoop MJ, Ab G. Import of proteins into peroxisomes and other microbodies. Biochem J. 1992 Sep 15;286(Pt 3):657–669. [PMC free article] [PubMed]
  • Gestetner B, Assa Y, Henis Y, Birk Y, Bondi A. Lucerne saponins. IV. Relationship between their chemical constitution, and haemolytic and antifungal activities. J Sci Food Agric. 1971 Apr;22(4):168–172. [PubMed]
  • Glover JR, Andrews DW, Rachubinski RA. Saccharomyces cerevisiae peroxisomal thiolase is imported as a dimer. Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10541–10545. [PMC free article] [PubMed]
  • Gould SG, Keller GA, Subramani S. Identification of a peroxisomal targeting signal at the carboxy terminus of firefly luciferase. J Cell Biol. 1987 Dec;105(6 Pt 2):2923–2931. [PMC free article] [PubMed]
  • Graham IA, Denby KJ, Leaver CJ. Carbon Catabolite Repression Regulates Glyoxylate Cycle Gene Expression in Cucumber. Plant Cell. 1994 May;6(5):761–772. [PMC free article] [PubMed]
  • Häusler T, Stierhof YD, Wirtz E, Clayton C. Import of a DHFR hybrid protein into glycosomes in vivo is not inhibited by the folate-analogue aminopterin. J Cell Biol. 1996 Feb;132(3):311–324. [PMC free article] [PubMed]
  • Hicks GR, Smith HM, Lobreaux S, Raikhel NV. Nuclear import in permeabilized protoplasts from higher plants has unique features. Plant Cell. 1996 Aug;8(8):1337–1352. [PMC free article] [PubMed]
  • Janssen BJ. A cDNA clone for isocitrate lyase from tomato. Plant Physiol. 1995 Jul;108(3):1339–1339. [PMC free article] [PubMed]
  • Kaufman RJ. Vectors used for expression in mammalian cells. Methods Enzymol. 1990;185:487–511. [PubMed]
  • Kolodziej PA, Young RA. Epitope tagging and protein surveillance. Methods Enzymol. 1991;194:508–519. [PubMed]
  • Kunce CM, Trelease RN, Turley RB. Purification and biosynthesis of cottonseed (Gossypium hirsutum L.) catalase. Biochem J. 1988 Apr 1;251(1):147–155. [PMC free article] [PubMed]
  • Marrison JL, Onyeocha I, Baker A, Leech RM. Recognition of Peroxisomes by Immunofluorescence in Transformed and Untransformed Tobacco Cells. Plant Physiol. 1993 Dec;103(4):1055–1059. [PMC free article] [PubMed]
  • McNew JA, Goodman JM. An oligomeric protein is imported into peroxisomes in vivo. J Cell Biol. 1994 Dec;127(5):1245–1257. [PMC free article] [PubMed]
  • McNew JA, Goodman JM. The targeting and assembly of peroxisomal proteins: some old rules do not apply. Trends Biochem Sci. 1996 Feb;21(2):54–58. [PubMed]
  • Olsen LJ, Ettinger WF, Damsz B, Matsudaira K, Webb MA, Harada JJ. Targeting of glyoxysomal proteins to peroxisomes in leaves and roots of a higher plant. Plant Cell. 1993 Aug;5(8):941–952. [PMC free article] [PubMed]
  • Onyeocha I, Behari R, Hill D, Baker A. Targeting of castor bean glyoxysomal isocitrate lyase to tobacco leaf peroxisomes. Plant Mol Biol. 1993 Jun;22(3):385–396. [PubMed]
  • Osumi T, Tsukamoto T, Hata S, Yokota S, Miura S, Fujiki Y, Hijikata M, Miyazawa S, Hashimoto T. Amino-terminal presequence of the precursor of peroxisomal 3-ketoacyl-CoA thiolase is a cleavable signal peptide for peroxisomal targeting. Biochem Biophys Res Commun. 1991 Dec 31;181(3):947–954. [PubMed]
  • Rachubinski RA, Subramani S. How proteins penetrate peroxisomes. Cell. 1995 Nov 17;83(4):525–528. [PubMed]
  • Rapp S, Soto U, Just WW. Import of firefly luciferase into peroxisomes of permeabilized Chinese hamster ovary cells: a model system to study peroxisomal protein import in vitro. Exp Cell Res. 1993 Mar;205(1):59–65. [PubMed]
  • Restrepo MA, Freed DD, Carrington JC. Nuclear transport of plant potyviral proteins. Plant Cell. 1990 Oct;2(10):987–998. [PMC free article] [PubMed]
  • Reynolds SJ, Smith SM. The isocitrate lyase gene of cucumber: isolation, characterisation and expression in cotyledons following seed germination. Plant Mol Biol. 1995 Feb;27(3):487–497. [PubMed]
  • Subramani S. Protein import into peroxisomes and biogenesis of the organelle. Annu Rev Cell Biol. 1993;9:445–478. [PubMed]
  • Subramani S. Convergence of model systems for peroxisome biogenesis. Curr Opin Cell Biol. 1996 Aug;8(4):513–518. [PubMed]
  • Swinkels BW, Gould SJ, Bodnar AG, Rachubinski RA, Subramani S. A novel, cleavable peroxisomal targeting signal at the amino-terminus of the rat 3-ketoacyl-CoA thiolase. EMBO J. 1991 Nov;10(11):3255–3262. [PMC free article] [PubMed]
  • Taylor KM, Kaplan CP, Gao X, Baker A. Localization and targeting of isocitrate lyases in Saccharomyces cerevisiae. Biochem J. 1996 Oct 1;319(Pt 1):255–262. [PMC free article] [PubMed]
  • Trelease RN, Hermerath CA, Turley RB, Kunce CM. Cottonseed malate synthase : purification and immunochemical characterization. Plant Physiol. 1987 Aug;84(4):1343–1349. [PMC free article] [PubMed]
  • Trelease RN, Choe SM, Jacobs BL. Conservative amino acid substitutions of the C-terminal tripeptide (Ala-Arg-Met) on cottonseed isocitrate lyase preserve import in vivo into mammalian cell peroxisomes. Eur J Cell Biol. 1994 Dec;65(2):269–279. [PubMed]
  • Trelease RN, Xie W, Lee MS, Mullen RT. Rat liver catalase is sorted to peroxisomes by its C-terminal tripeptide Ala-Asn-Leu, not by the internal Ser-Lys-Leu motif. Eur J Cell Biol. 1996 Nov;71(3):248–258. [PubMed]
  • Turley RB, Choe SM, Ni W, Trelease RN. Nucleotide sequence of cottonseed malate synthase. Nucleic Acids Res. 1990 Jun 25;18(12):3643–3643. [PMC free article] [PubMed]
  • Vanni P, Giachetti E, Pinzauti G, McFadden BA. Comparative structure, function and regulation of isocitrate lyase, an important assimilatory enzyme. Comp Biochem Physiol B. 1990;95(3):431–458. [PubMed]
  • Walton PA, Hill PE, Subramani S. Import of stably folded proteins into peroxisomes. Mol Biol Cell. 1995 Jun;6(6):675–683. [PMC free article] [PubMed]
  • Wendland M, Subramani S. Cytosol-dependent peroxisomal protein import in a permeabilized cell system. J Cell Biol. 1993 Feb;120(3):675–685. [PMC free article] [PubMed]
  • Wigler M, Sweet R, Sim GK, Wold B, Pellicer A, Lacy E, Maniatis T, Silverstein S, Axel R. Transformation of mammalian cells with genes from procaryotes and eucaryotes. Cell. 1979 Apr;16(4):777–785. [PubMed]
  • Wolvetang EJ, Tager JM, Wanders RJ. Latency of the peroxisomal enzyme acyl-CoA:dihydroxyacetonephosphate acyltransferase in digitonin-permeabilized fibroblasts: the effect of ATP and ATPase inhibitors. Biochem Biophys Res Commun. 1990 Aug 16;170(3):1135–1143. [PubMed]

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