Abstract
We have examined transcription in an early diverging eukaryote by analyzing the effect of the fungus-derived toxin α-amanitin on the transcription of protein-coding genes of the protistTrichomonas vaginalis. In contrast to that typical in eukaryotes, the RNA polymerase that transcribesT. vaginalis protein-coding genes is relatively resistant to α-amanitin (50% inhibition = 250 μg α-amanitin/ml). We have also characterized the gene encoding the largest subunit of RNA polymerase II, the subunit that binds α-amanitin. This protein is 41% identical to the mouse RNA polymerase II. Sequence analysis of the 50-amino-acid region thought to bind α-amanitin shows that this region of the trichomonad RNA polymerase II lacks many of the conserved amino acids present in the putative binding site, in agreement with the observed insensitivity to this inhibitor. Similar to other RNA polymerase Its analyzed from ancient eukaryotes, theT. vaginalis RNA polymerase II lacks the typical heptapeptide (Tyr-Ser-Pro-Thr-Ser-Pro-Ser) repeat carboxyl-terminal domain (CTD) that is a hallmark of higher eukaryotic RNA polymerase IIs. The trichomonad enzyme, however, does contain a short modified CTD that is rich in the amino acid residues that compose the repeat. These data suggest thatT. vaginalis protein-coding genes are transcribed by a RNA polymerase II that is relatively insensitive to α-amanitin and that differs from typical eukaryotic RNA polymerase Its as it lacks a heptapeptide repeated CTD.
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Allison LA, Moyle M, Shales M, Ingles CJ (1985) Extensive homology among the largest subunits of eukaryotic and prokaryotic RNA polymerases. Cell 42:599–610
Ahearn JM, Bartolomei MS, West ML, Cisek LJ, Corden JL (1987) Cloning and sequence analysis of the mouse genomic locus encoding the largest subunit of RNA polymerase II. J Biol Chem 262: 10695–10705
Bartolomei MS, Corden JL (1987) Localization of an α-amanitin resistance mutation in the gene encoding the largest subunit of mouse RNA polymerase II. Mol Cell Biol 7:586–594
Biggs J, Searles LL, Greenleaf AL (1985) Structure of the eukaryotic transcription apparatus: features of the gene for the largest subunit ofDrosophila RNA polymerase II. Cell 42:611–621
Bird DM, Riddle DL (1989) Molecular cloning and sequencing of ama-I, the gene encoding the largest subunit ofCaenorhabditis elegans RNA polymerase II. Mol Cell Biol 9:4119–4130
Cadena D, Dahmus M (1987) Messenger RNA synthesis in mammalian cells is catalyzed by phosphorylated form of RNA polymerase II. J Biol Chem 262:12468–12474
Chen Y, Weeks J, Mortin MA, Greenleaf AL (1993) Mapping mutations in genes encoding the two large subunits ofDrosophila RNA polymerase II defines domains essential for basic transcription functions and for proper expression of developmental genes. Mol Cell Biol 13:4214–4222
Chung HM, Lee MGS, van der Ploeg LHT (1993) RNA polymerase I-mediated protein-coding gene expression inTrypanosoma brucei. Parasitol Today 9:17–17
Corden JL (1990) Tails of RNA polymerase II. Trends Biochem Sci 15:383–387
Corden JL, Cadena DL, Ahearn J Jr, Dahmus ME (1985) A unique structure at the carboxyl terminus of the largest subunit of eukaryotic RNA polymerase II. Proc Natl Acad Sci USA 82:7934–7938
Cornelissen AWCA, Evers R, Kock J (1988) Structure and sequence of genes encoding subunits of eukaryotic RNA polymerases. Oxf Surv Eukaryot Genes 5:91–131
Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12: 387–395
Edman U, Meraz MA, Rausser S, Agabian N, Meza I (1990) Characterization of an immunodominant variable surface antigen from pathogenic and nonpathogenicEntamoeba histolyica. J Exp Med 172:879–888
Evers R, Hammer A, Cornelissen AWCA (1989a) Unusual C-terminal domain of the largest subunit of RNA polymerase II ofCrithidia fasciculata. Nucleic Acids Res 17:3403–3413
Evers R, Hammer A, Kock J, Jess W, Borst P, Memets S, Cornelissen AWCA (1989b)Trypanosoma brucei contains two RNA polymerase II largest subunit genes with an altered C-terminal domain. Cell 56:585–597
Ey PL, Khanna K, Manning PA, Mayrhofer G (1993) The gene encoding a 69-kilodalton major surface protein ofGiardia intestinalis trophozoites. Mol Biochem Parasitol 58:247–257
Felsenstein J (1993) PHYLIP users manual version 3.5c. University of Washington, Seattle
Gorrell TE, Yarlett N, Müller M (1984) Isolation and characterization ofTrichomonas vaginalis ferredoxin. Carlsberg Res Commun 49: 259–268
Greenleaf AL (1993) A positive addition to a negative tail's tale. Proc Natl Acad Sci USA 90:10896–10897
Janz L, Clayton C (1994) The PARP and rRNA promoters ofTrypanosoma brucei are composed of dissimilar sequence elements that are functionally interchangeable. Mol Cell Biol 14:5804–5811
Jess W, Hammer A, Comelissen AWCA (1989) Complete sequence of the gene encoding the largest subunit of RNA polymerase I ofTrypanosome brucei. FEBS Lett 249:123–128
Jokerst RS, Weeks JR, Zehring WA, Greenleaf AL (1989) Analysis of the gene encoding the largest subunit of RNA polymerase II inDrosophilia. Mol Gen Genet 215:266–275
Klenk H-P, Zillig W, Lanzendörfer M, Grampp B, Palm P (1995) Location of protist lineages in a phylogenetie tree inferred from sequences of DNA-dependent RNA polymerases. Archiv Protisten Kunde 145:221–230
Kock J, Evers R, Cornelissen AWCA (1988) Structure and sequence of the gene for the largest subunit of trypanosomal RNA polymerase III. Nucleic Acids Res 16:8753–8772
Kooter JM, Borst P (1984) α-Amanitin-insensitive transcription of variant surface glycoprotein genes provides further evidence for discontinuous transcription in trypanosomes. Nucleic Acids Res 12:9457–9472
la Cour TFM, Nyborg J, Thirup S, Clark BFC (1985) Structural details of the binding of guanosine diphosphate to elongation factor Tu fromE. coli as studies by X-ray crystallography. EMBO J 4:2385–2388
Lahti CJ, Bradley PJ, Johnson PJ (1994) Molecular characterization of the α-subunit ofTrichomonas vaginalis hydrogenosomal succinyl COA synthetase. Mol Biochem Parasitol 66:309–318
Leffers H, Gropp F, Lottspeich F, Zillig W, Garrett RA (1989) Sequence, organization, transcription and evolution of RNA polymerase subunit genes from the archaebacterial extreme halophilesHalobacterium halobium and Halococcus morrhuae. J Mol Biol 206: 1–17
Leipe DD, Gunderson JH, Nerad TA, Sogin ML (1993) Small subunit ribosomal RNA+ ofHexamita inflata and the quest for the first branch in the eukaryotic tree. Mol Biochem Parasitol 59:41–48
Li WB, Bzik DJ, Gu H, Tanaka M (1989) An enlarged largest subunit ofPlasmodium falciparum RNA polymerase II defines conserved and variable RNA polymerase domains. Nucleic Acids Res 17: 9621–9636
Li W-B, Bzik DJ, Tanaka M, Gu H, Fox BA, Inselburg J (1991) Characterization of the gene encoding the largest subunit ofPlasmodium falciparum RNA polymerase III. Mol Biochem Parasitol 46:229–240
Lioutas C, Tarnich E (1995) Transcription of protein coding genes in Entamoeba histolytica is insensitive to high concentrations of α-amanitin. Mol Biochem Parasitol 73:259–261
McKnight SL, Kingsbury R (1982) Transcriptional control signals of a eukaryotic protein-coding gene. Science 217:316–324
Mémet S, Gouy M, Marck C, Sentenac A, Buhler J-M (1988) RPA190, the gene encoding for the largest subunit of yeast RNA polymerase A. J Biol Chem 263:2830–2839
Müller M (1988) Energy metabolism of protozoa without mitochondria. Annu Rev Microbiol 42:465–488
Müller M (1993) The hydrogenosome. J Gen Microbiol 139:2879–2889
Ovchinnikov YA, Monastyrskaya GS, Gubanov VV, Gureyev SO, Salomatina IS, Shuvaea TM, Lipkin VM, Svederlov ED (1982) The primary structure ofE. coli RNA polymerase. Nucleotide sequence of the rpoC gene and amino acid sequence of the β′-subunit. Nucleic Acids Res 10:4035–4044
Pühler G, Lottspeich F, Zillig W (1989) Organization and nucleotide sequence of the genes encoding the large subunits A, B and C of the DNA-dependent RNA polymerase of the archaebacteriumSulfolobus acidocaldarius. Nucleic Acids Res 17:4517–4534
Quon DVK, d'Oliveira CE, Johnson PJ (1992) Reduced transcription of the ferredoxin gene in metronidazole-resistantTrichomonas vaginalis. Proc Natl Acad Sci USA 89:4402–4406
Quon DVK, Delgadillo MG, Khachi A, Smale ST, Johnson PJ (1994) Similarity between a ubiquitous promoter element in an ancient eukaryote and mammalian initiator elements. Proc Natl Acad Sci USA 91:4579–4583
Rudenko G, Le Blancq S, Smith J, Lee MG, Rattray A, van der Ploeg LHT (1990) Procyclic acidic repetitive protein (PARP) genes located in an unusually small alpha-amanitin-resistant transcription unit—PARP promoter activity assayed by transient DNA transfection ofTrypanosoma brucei. Mol Cell Biol 10:3492–3504
Rudenko G, Lee MGS, van der Ploeg LHT (1992) The PARP and VSG genes ofTrypanosoma brucei do not resemble RNA polymerase-II transcription units in sensitivity to sarkosyl in nuclear run-on assays. Nucleic Acids Res 20:303–306
Sanford T, Golomb M, Riddle DL (1983) RNA polymerase II from wild type and alpha-amanitin-resistant strains ofCaenorhabditis elegans. J. Biol Chem 258:12804–12809
Sawadogo M, Sentenac A (1990) RNA polymerase B (II) and general transcription factors. Annu Rev Biochem 59:711–754
Sentenac A (1985) Eukaryotic RNA polymerases. Crit Rev Biochem 18:31–90
Smale ST (1994) Core promoter architecture for eukaryotic protein-coding genes. In: Conaway RC, Conaway JW, eds. Transcription: mechanisms and regulation. Raven Press, New York, pp 63–81
Smith JL, Levin JR, Ingles CJ, Agabian N (1989) In trypanosomes the homolog of the largest subunit of RNA polymerase II is incoded by two genes and has a highly unusual C-terminal domain structure. Cell 56:815–827
Stunnenberg HG, Wennekes LMJ, Spierings T, van den Broek HWJ (1981) An α-Amanitin-resistant DNA-dependent RNA polymerase II from the fungusAspergillus nidulans. Eur J Biochem 117:121–129
Wang AL, Wang CC (1985) A linear double-stranded RNA inTrichomonas vaginalis. J Biol Chem 260:3697–3702
Wieland T, Faulstich J (1978) Amatoxins, phallotoxins, phallolysin, and antmanide: the biologically active components of poisonousAmanita mushrooms. Crit Rev Biochem 5:185–260
Wintzerith M, Acker J, Vicaire S, Vigneron M, Kedinger C (1992) Complete sequence of the human RNA polymerase II largest subunit. Nucleic Acids Res 20:910
Young RA (1991) RNA polymerase II. Anna Rev Biochem 60:689–715
Zhang J, Corden JL (1991) Identification of phosphorylation sites in the repetitive carboxyl-terminal domain of the mouse RNA polymerase II largest subunit. J Biol Chem 266:2290–2296
Zomerdijk JCBM, Kieft R, Shiels PG, Borst P (1991) Alpha-amanitin-resistant transcription units in trypanosomes: a comparison of promoter sequences for a VSG gene expression site and for the ribosomal RNA genes. Nucleic Acids Res 19:5153–5158
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Quon, D.V.K., Delgadillo, M.G. & Johnson, P.J. Transcription in the early diverging eukaryoteTrichomonas vaginalis: An unusual RNA polymerase II and α-amanitin-resistant transcription of protein-coding genes. J Mol Evol 43, 253–262 (1996). https://doi.org/10.1007/BF02338833
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DOI: https://doi.org/10.1007/BF02338833