We have identified and cloned four trypanosomal RNA polymerase largest subunit genes. Here, we pr... more We have identified and cloned four trypanosomal RNA polymerase largest subunit genes. Here, we present the molecular analysis of two genes, Trp4.8 and Trp5.9. The sequence of these genes shows that they are almost identical to each other and indicates that they encode the largest subunit of RNA polymerase II. Both genes contain a C-terminal extension that is clearly distinct from that of other eukaryotic RNA polymerase II genes, because it lacks the common tandemly repeated heptapeptide sequence and is rich in acidic amino acids. It shares many potential phosphorylation sites, however, with the C-terminal extension of other eukaryotic RNA polymerase II large subunits. The presence of two RNA polymerase II loci suggests that a fourth RNA polymerase could be formed. Interestingly, the fourth gene is only found in species exhibiting antigenic variation.
We have set out to clone the trypanosomal gene encoding the largest subunit of RNA polymerase I. ... more We have set out to clone the trypanosomal gene encoding the largest subunit of RNA polymerase I. We screened a genomic library with a synthetic oligonucleotide probe encoding an eleven amino acid sequence motif, YNAD~DEMN, which has been found in all eukaryotic RNA polymerase largest subunit genes analyzed so far. We isolated the Trpl 1 locus and determined the complete sequence of the gene encoded within this locus. The deduced amino acid sequence contains the highly conserved RNA polymerase domains as well as the previously identified RNA polymerase I-specific hydrophilic insertions. Therefore, the gene most closely resembles the largest subunit of RNA polymerase I.
We have sequenced the genes encoding to largest subunits of the three classes of DNA-dependent RN... more We have sequenced the genes encoding to largest subunits of the three classes of DNA-dependent RNA polymerases of Trypanosoma brucei. The nucleotide and deduced amino acid sequences were compared and aligned with the corresponding sequences of other eukaryotes. Phylogenetic relationships were subsequently calculated with a distant matrix, a bootstrapped parsimony and a maximum-likelihood method. These independent calculations resulted in trees with very similar topologies. The analyses show that all the largest subunits of T. brucei are evolutionarily distant members within each of the three RNA polymerase classes. An early separation of the trypanosomal subunits from the eukaryotic lineage might form the fundamental basis for the unusual transcription process of this species. Finally, all dendrograms show a separate ramification for the largest subunit of RNA polymerase I, II and III. RNA polymerase II and/or III form a bifurcation with the archaebacterial lineage, RNA polymerase I, however, arises separately from the eubacterial beta' lineage. This suggests that the three eukaryotic RNA polymerase classes are not simply derived by two gene duplications of an ancestral gene with subsequent differentiation.
We have identified and cloned four trypanosomal RNA polymerase largest subunit genes. Here, we pr... more We have identified and cloned four trypanosomal RNA polymerase largest subunit genes. Here, we present the molecular analysis of two genes, Trp4.8 and Trp5.9. The sequence of these genes shows that they are almost identical to each other and indicates that they encode the largest subunit of RNA polymerase II. Both genes contain a C-terminal extension that is clearly distinct from that of other eukaryotic RNA polymerase II genes, because it lacks the common tandemly repeated heptapeptide sequence and is rich in acidic amino acids. It shares many potential phosphorylation sites, however, with the C-terminal extension of other eukaryotic RNA polymerase II large subunits. The presence of two RNA polymerase II loci suggests that a fourth RNA polymerase could be formed. Interestingly, the fourth gene is only found in species exhibiting antigenic variation.
We have set out to clone the trypanosomal gene encoding the largest subunit of RNA polymerase I. ... more We have set out to clone the trypanosomal gene encoding the largest subunit of RNA polymerase I. We screened a genomic library with a synthetic oligonucleotide probe encoding an eleven amino acid sequence motif, YNAD~DEMN, which has been found in all eukaryotic RNA polymerase largest subunit genes analyzed so far. We isolated the Trpl 1 locus and determined the complete sequence of the gene encoded within this locus. The deduced amino acid sequence contains the highly conserved RNA polymerase domains as well as the previously identified RNA polymerase I-specific hydrophilic insertions. Therefore, the gene most closely resembles the largest subunit of RNA polymerase I.
We have sequenced the genes encoding to largest subunits of the three classes of DNA-dependent RN... more We have sequenced the genes encoding to largest subunits of the three classes of DNA-dependent RNA polymerases of Trypanosoma brucei. The nucleotide and deduced amino acid sequences were compared and aligned with the corresponding sequences of other eukaryotes. Phylogenetic relationships were subsequently calculated with a distant matrix, a bootstrapped parsimony and a maximum-likelihood method. These independent calculations resulted in trees with very similar topologies. The analyses show that all the largest subunits of T. brucei are evolutionarily distant members within each of the three RNA polymerase classes. An early separation of the trypanosomal subunits from the eukaryotic lineage might form the fundamental basis for the unusual transcription process of this species. Finally, all dendrograms show a separate ramification for the largest subunit of RNA polymerase I, II and III. RNA polymerase II and/or III form a bifurcation with the archaebacterial lineage, RNA polymerase I, however, arises separately from the eubacterial beta' lineage. This suggests that the three eukaryotic RNA polymerase classes are not simply derived by two gene duplications of an ancestral gene with subsequent differentiation.
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Papers by Waldemar Jess