Transcription in the early diverging eukaryote Trichomonas vaginalis: an unusual RNA polymerase II and alpha-amanitin-resistant transcription of protein-coding genes.
We have examined transcription in an early diverging eukaryote by analyzing the effect of the fungus-derived toxin alpha-amanitin on the transcription of protein-coding genes of the protist Trichomonas vaginalis. In contrast to that typical in eukaryotes, the RNA polymerase that transcribes T. vaginalis protein-coding genes is relatively resistant to alpha-amanitin (50% inhibition = 250 microg alpha-amanitin/ml). We have also characterized the gene encoding the largest subunit of RNA polymerase II, the subunit that binds alpha-amanitin. This protein is 41% identical to the mouse RNA polymerase II. Sequence analysis of the 50-amino-acid region thought to bind alpha-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 IIs analyzed from ancient eukaryotes, the T. 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 that T. vaginalis protein-coding genes are transcribed by a RNA polymerase II that is relatively insensitive to alpha-amanitin and that differs from typical eukaryotic RNA polymerase IIs as it lacks a heptapeptide repeated CTD.
Quon
D V
DV
Department of Microbiology and Immunology, School of Medicine, University of California, Los Angeles, 90095-1747, USA.
Delgadillo
M G
MG
Johnson
P J
PJ
D000546:Amanitins / Q000494:pharmacology*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D016208:Databases, Factual; D004351:Drug Resistance / Q000235:genetics*; D017125:Genes, Protozoan; D014644:Genetic Variation; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D015800:Protozoan Proteins / Q000096:biosynthesis* / Q000235:genetics; D012318:RNA Polymerase I / Q000737:chemistry / Q000235:genetics; D012319:RNA Polymerase II / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D012320:RNA Polymerase III / Q000737:chemistry / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D012984:Software; D014158:Transcription, Genetic; D014246:Trichomonas vaginalis / Q000235:genetics*
D000546:Amanitins; D017931:DNA Primers; D015800:Protozoan Proteins; D012319:RNA Polymerase II; D012318:RNA Polymerase I; D012320:RNA Polymerase III