Abstract
In Trypanoplasma borelli, a representative of the Bodonina within the Kinetoplastida, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) activity was detected in both the cytosol and glycosomes. This situation is similar to that previously found in Trypanosomatidae, belonging to a different Kinetoplastida suborder. In Trypanosomatidae different isoenzymes, only distantly related, are responsible for the activity in the two cell compartments. In contrast, immunoblot analysis indicated that the GAPDH activity in cytosol and glycosomes of T. borelli should be attributed to identical or at least very similar proteins related to the glycosomal GAPDH of Trypanosomatidae. Moreover, only genes related to the glycosomal GAPDH genes of Trypanosomatidae could be detected. All attempts to identify a gene related to the one coding for the trypanosomatid cytosolic GAPDH remained unsuccessful. Two tandemly arranged genes were found which are 95% identical. The two encoded polypeptides differ in 17 residues. Their sequences are 72–77% identical to the glycosomal GAPDH of the other Kinetoplastida and share with them some characteristic features: an excess of positively charged residues, specific insertions, and a small carboxy-terminal extension containing the sequence -AKL. This tripeptide conforms to the consensus signal for targeting of proteins to glycosomes. One of the two gene copies has undergone some mutations at positions coding for highly conserved residues of the active site and the NAD+-binding domain of GAPDH. Modeling of the protein's three-dimensional structure suggested that several of the substitutions compensate each other, retaining the functional coenzyme-binding capacity, although this binding may be less tight. The presented analysis of GAPDH in T. borelli gives further support to the assertion that one isoenzyme, the cytosolic one, was acquired by horizontal gene transfer during the evolution of the Kinetoplastida, in the lineage leading to the suborder Trypanosomatina (Trypanosome, Leishmania), after the divergence from the Bodonina (Trypanoplasma). Furthermore, the data clearly suggest that the original GAPDH of the Kinetoplastida has been compartmentalized during evolution.
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Abbreviations
- GAPDH:
-
glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.12)
- HK:
-
hexokinase (EC 2.7.1.1)
- PGI:
-
glucosephosphate isomerase (EC 5.3.1.9)
- PGK:
-
phosphoglycerate kinase (EC 2.7.2.3)
- PYK:
-
pyruvate kinase (EC 2.7.1.40)
- TIM:
-
triosephosphate isomerase (EC 5.3.1.1)
- SDS:
-
sodium dodecyl sulfate
- SSC:
-
saline sodium citrate (0.15 M NaCl, 15 mM sodium citrate, pH 7.0)
- MYR:
-
millions of years
References
Alefounder PR, Perham RN (1989) Identification, molecular cloning and sequence analysis of a gene cluster encoding the Class II fructose 1,6-bisphosphate aldolase, 3-phosphoglycerate kinase and a putative second glyceraldehyde 3-phosphate dehydrogenase of Escherichia coli. Mol Microbiol 3:723–732
Alexander K, Parsons M (1991) A phosphoglycerate kinase-like molecule localized to glycosomal microbodies: evidence that the topogenic signal is not at the C-terminus. Mol Biochem Parasitol 46: 1–10
Blattner J, Swinkels BW, Dorsam H, Prospero T, Subramani S, Clayton C (1992) Glycosome assembly in trypanosomes: variations in the acceptable degeneracy of a COOH-terminal microbody targeting signal. J Cell Biol 119:1129–1136
Borst P, Swinkels BW (1989) The evolutionary origin of glycosomes; how glycolysis moved from cytosol to organelle. In: Grunberg-Manago M, Clark BFC, Zachau HG (eds) Evolutionary tinkering in gene expression. Plenum Publishing Corporation, pp 163–174
Branlant G, Branlant C (1985) Nucleotide sequence of the Escherichia coli gap gene. Different evolutionary behaviour of the NAD+-binding domain and of the catalytic domain of D-glyceraldehyde-3-phosphate dehydrogenase. Eur J Biochem 150:61–66
Brun R, Schönenberger M (1979) Cultivation and in vitro cloning of procyclic culture forms of Trypanosoma brucei in a semi-defined medium. Acta Trop (Basel) 36:289–292
Callens M, Kuntz DA, Opperdoes FR (1991) Characterization of pyruvate kinase from Trypanosoma brucei and its role in the regulation of carbohydrate metabolism. Mol Biochem Parasitol 47:19–30
Cavalier-Smith T (1990) Symbiotic origin of peroxisomes. In: Nardon P, Gianinazii-Pecurson V, Grenier AM, Margulis L, Smith DC (eds) Endocytobiology IV. pp 515–520
Chen J-H, Gibson JL, McCue LA, Tabita FR (1991) Identification, expression, and deduced primary structure of transketolase and other enzymes encoded within the form II carbon dioxide fixation operon of Rhodobacter sphaeroides. J Biol Chem 266: 20447–20452
Conway T, Sewell GW, Ingram LO (1987) Glyceraldehyde-3-phosphate dehydrogenase gene from Zymomonas mobilis: cloning, sequencing, and identification of promoter region. J Bacteriol 169: 5653–5662
Doolittle RF, Feng DF, Anderson KL, Alberro MR (1990) A naturally occurring horizontal gene transfer from a eukaryote to a prokaryote. J Mol Evol 31:383–388
Efron B (1982) The jack-knife, the bootstrap and other resampling plans. Society for Industrial and Applied Mathematics, Philadelphia
Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791
Felsenstein J (1993) PHYLIP manual, version 3.5
Fitch WM, Margoliash E (1967) Construction of phylogenetic trees. Science 155:279–284
Fernandez AP, Nelson K, Beverley SM (1993) Evolution of nuclear ribosomal RNAs in kinetoplastid protozoa: perspectives on the age and origins of parasitism. Proc Natl Acad Sci USA 90:11608–11612
Fothergill-Gilmore LA, Michels PAM (1993) Evolution of glycolysis. Progr Biophys Mol Biol 59:105–235
Gilbert DG (1993) ReadSeq manual
Goudot-Crozel V, Caillol D, Djabali M, Dessein AJ (1989) The major parasite surface antigen associated with human resistance to schistosomiasis is a 37 kD glyceraldehyde-3P-dehydrogenase. J Exp Med 170:2065–2080
Gould SJ, Keller GA, Hosken N, Wilkinson J, Subramani S (1989) A conserved tripeptide sorts proteins to peroxisomes. J Cell Biol 108: 1657–1664
Harris JI, Perham RN (1968) Glyceraldehyde 3-phosphate dehydrogenase from pig muscle. Nature 219:1025–1028
Hannaert V, Blaauw M, Kohl L, Allert S, Opperdoes FR, Michels PAM (1992) Molecular analysis of the cytosolic and glycosomal glyceraldehyde-3-phosphate dehydrogenase in Leishmania mexicana. Mol Biochem Parasitol 55:115–126
Hannaert V, Michels PAM (1994) Structure, function and biogenesis of glycosomes in Kinetoplastida. J Bioenerg Biomembr 20:205–212
Hart DT, Opperdoes FR (1984) The occurrence of glycosomes (microbodies) in the promastigote stage of four major Leishmania species. Mol Biochem Parasitol 13:159–172
Hasegawa M, Fujiwara M (1993) Relative efficiencies of the maximum likelihood, maximum parsimony, and neighbor-joining methods for estimating protein phylogeny. Mol Phylogenet Evol 2:1–5
Hecht RM, Garza A, Lee Y-H, Miller MD, Pisegna MA (1989) Nucleotide sequence of the glyceraldehyde-3-phosphate dehydrogenase gene from Thermus aquaticus YT1. Nucleic Acids Res 17:10–23
Holland JP, Holland MJ (1979) The primary structure of a glyceraldehyde-3-phosphate dehydrogenase gene from Saccharomyces cerevisiae. J Biol Chem 254:9839–9845
Keegan FP, Blum JJ (1993) Incorporation of label from acetate and laurate into mannan of Leishmania donovani via the glyoxylate cycle. J Euk Microbiol 40:730–732
Kendall G, Wilderspin AWF, Ashall F, Miles MA, Kelly JM (1990) Trypanosoma cruzi glycosomal glyceraldehyde-3-phosphate dehydrogenase does not conform to the “hotspot” model of topogenesis. EMBO J 9:2751–2758
Kimura M (1983) The neutral theory of molecular evolution. Cambridge University Press, Cambridge.
Kohl L, Callens M, Wierenga RK, Opperdoes FR, Michels PAM (1994) Triosephosphate isomerase of Leishmania mexicana mexicana. Cloning and characterization of the gene, overexpression in Escherichia coli and analysis of the protein. Eur J Biochem 220: 331–338
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Lambeir A-M, Loiseau AM, Kuntz DA, Vellieux FM, Michels PAM, Opperdoes FR (1991) The cytosolic and glycosomal glyceraldehyde-3-phosphate dehydrogenase from T. brucei: kinetic properties and comparison with homologous enzymes. Eur J Biochem 198: 429–435
Le Blancq SM, Swinkels BW, Gibson WC, Borst P (1988) Evidence for gene conversion between the phosphoglycerate kinase genes of Trypanosoma brucei. J Mol Biol 200:439–447
Marchand M, Kooystra U, Wierenga RK, Lambeir A-M, Van Beeumen J, Opperdoes FR, Michels PAM (1989) Glucosephosphate isomerase from Trypanosoma brucei. Cloning and characterization of the gene and analysis of the enzyme. Eur J Biochem 184:455–464
Markos A, Miretsky A, Müller M (1993) A glyceraldehyde-3-phosphate dehydrogenase with eubacterial features in the amitochondriate eukaryote Trichomonas vaginalis. J Mol Evol 37:631–643
Martin W, Cerff R (1986) Prokaryotic features of a nucleus-encoded enzyme. cDNA sequences for chloroplast and cytosolic glyceraldehyde-3-phosphate dehydrogenases from mustard (Sinapis alba). Eur J Biochem 159:323–331
Martin W, Brinkmann H, Savona C, Cerff R (1993) Evidence for a chimaeric nature of nuclear genomes: eubacterial origin of eukaryotic glyceraldehyde-3-phosphate dehydrogenase genes. Proc Natl Acad Sci USA 90:8692–8696
Martinez P, Martin W, Cerff R (1989) Structure, evolution and anaerobic regulation of a nuclear gene encoding cytosolic glyceraldehyde-3-phosphate dehydrogenase from maize. J Mol Biol 208:551–565
McLaughlin PJ, Dayhoff MO (1973) Eukaryotic evolution: a view based on cyrochrome C sequence data. J Mol Evol 2:99–116
Michels PAM, Poliszczak A, Osinga KA, Misset O, Van Beeumen J, Wieringa RK, Borst P, Opperdoes FR (1986) Two tandemly linked genes code for the glycosomal glyceraldehyde-phosphate dehydrogenase in Trypanosoma brucei. EMBO J 5:1049–1056
Michels PAM, Marchand M, Kohl L, Allert S, Wierenga RK, Opperdoes FR (1991) The cytosolic and glycosomal isoenzymes of glyceraldehyde-3-phosphate dehydrogenase in Trypanosoma brucei have a distant evolutionary relationship. Eur J Biochem 198:421–428
Michels PAM, Opperdoes FR (1991) The evolutionary origin of glycosomes. Parasitol Today 7:105–109
Michels PAM, Hannaert V (1994) The evolution of kinetoplastid glycosomes. J Bioenerg Biomembr 26:213–219
Misset O, Opperdoes FR (1984) Simultaneous purification of hexokinase, class-I fructose-bisphosphate aldolase, triosephosphate isomerase and phosphoglycerate kinase from Trypanosoma brucei. Eur J Biochem 144:475–483
Misset O, Van Beeumen J, Lambeir AM, Van Der Meer R, Opperdoes FR (1987) Glyceraldehyde-phosphate dehydrogenase from Trypanosoma brucei: comparison of the glycosomal and cytosolic isoenzymes. Eur J Biochem 162:501–507
Opperdoes FR, Borst P (1977) Localization of nine glycolytic enzymes in a microbody-like organelle in Trypanosoma brucei: the glycosome. FEBS Lett 80:360–364
Opperdoes FR (1987) Compartmentation of carbohydrate metabolism in trypanosomes. Ann Rev Microbiol 41:127–151
Opperdoes FR, Nohynkova E, Van Schaftingen E, Lambeir A-M, Veenhuis M, Van Roy J (1988) Demonstration of glycosomes (microbodies) in the Bodonid flagellate Trypanoplasma borelli (Protozoa, Kinetoplastida). Mol Biochem Parasitol 30:155–164
Osinga KA, Swinkels BW, Gibson WC, Borst P, Veeneman GH, Van Boom JH, Michels PAM, Opperdoes FR (1985) Topogenesis of microbody enzymes: a sequence comparison of the genes for the glycosomal (microbody) and cytosolic phosphoglycerate kinases of Trypanosoma brucei. EMBO J 4:3811–3817
Punt P, Dingemanse MA, Jacobs-Meijsing BJM, Pouwels PH, Van den Hondel CAMJJ (1988) Isolation and characterization of the glyceraldehyde-3-phosphate dehydrogenase gene of Aspergillus nidulans. Gene 69:49–57
Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425
Schultes V, Deutzmann R, Jaenicke R (1990) Complete amino-acid sequence of glyceraldehyde-3-phosphate dehydrogenase from the hyperthermophilic eubacterium Thermotoga maritima. Eur J Biochem 192:25–31
Shih M-C, Heinrich PC, Goodman HM (1991) Cloning and chromosomal mapping of nuclear genes encoding chloroplast and cytosolic glyceraldehyde-3-phosphate dehydrogenase from Arabidopsis thaliana. Gene 104:133–138
Sogin ML, Elwood HJ, Gunderson JH (1986) Evolutionary diversity of eukaryotic small-subunit rRNA genes. Proc Natl Acad Sci USA 83:1383–1387
Sommer JM, Cheng Q-L, Keller GA, Wang CC (1992) In vivo import of firefly luciferase into the glycosomes of Trypanosoma brucei and mutational analysis of the C-terminal targeting signal. Mol Biol Cell 3:749–759
Stein S, Bohlen P, Stone J, Dairman W, Udenfriend S (1973) Amino-acid analysis with fluorescamine at the picomole level. Arch Biochem Biophys 155:203–212
Stone EM, Rothblum KN, Alevy MC, Koo TM, Schwartz RJ (1985) Complete sequence of the chicken glyceraldehyde-3-phosphate dehydrogenase gene. Proc Natl Acad Sci USA 82:1628–1642
Swinkels BW, Evers R, Borst P (1988) The topogenic signal of the glycosomal (microbody) phosphoglycerate kinase of Crithidia fasciculata resides in a carboxy-terminal extension. EMBO J 7:1159–1165
Swinkels BW, Loiseau A, Opperdoes FR, Borst P (1992) A phosphoglycerate kinase-related gene conserved between Trypanosoma brucei and Crithidia fasciculata. Mol Biochem Parasitol 50:69–78
Tso JY, Sun X-H, Kao T-H, Reece KS, Wu R (1985) Isolation and characterization of rat and human glyceraldehyde-3-phosphate dehydrogenase cDNAs: genomic complexity and molecular evolution of the gene. Nucleic Acids Res 13:2485–2502
Van der Ploeg LHT, Bernards A, Rijsewijk FAM, Borst P (1982) Characterization of the DNA duplication-transposition that controls the expression of two genes for variant surface glycoproteins in Trypanosoma brucei. Nucleic Acids Res 10:539–609
Vellieux FMD, Hajdu J, Verlinde CLMJ, Groendijk H, Read RJ, Greenhough TJ, Campbell JW, Kalk KH, Littlechild JA, Watson HC, Hol WGJ (1993) Structure of glycosomal glyceraldehyde-3-phosphate dehydrogenase from Trypanosoma brucei determined from Laue data. Proc Natl Acad Sci USA 90:2355–2359
Viaene A, Dhaese P (1989) Sequence of the glyceraldehyde-3-phosphate dehydrogenase gene from Bacillus subtilis. Nucleic Acids Res 17:12–51
Visser N, Opperdoes FR (1980) Glycolysis in Trypanosoma brucei. Eur J Biochem 103:623–632
Wierenga RK, Swinkels B, Michels PAM, Osinga K, Misset O, Van Beeumen J, Gibson WC, Postma JPM, Borst P, Opperdoes FR, Hol WGJ (1987) Common elements on the surface of glycolytic enzymes from Trypanosoma brucei may serve as topogenic signals for import into glycosomes. EMBO J 6:215–221
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Nucleotide sequence data reported in this paper have been submitted to the EMBL/Genbank/DDBJ nucleotide sequence databases under accession number X74535
Correspondence to: P.A.M. Michels
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Wiemer, E.A.C., Hannaert, V., van den Ijssel, P.R.L.A. et al. Molecular analysis of glyceraldehyde-3-phosphate dehydrogenase in Trypanoplasma borelli: An evolutionary scenario of subcellular compartmentation in Kinetoplastida. J Mol Evol 40, 443–454 (1995). https://doi.org/10.1007/BF00164030
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DOI: https://doi.org/10.1007/BF00164030