Structure and transcription of a P-ATPase gene from Trypanosoma brucei.
A putative ATPase gene was cloned from Trypanosoma brucei genomic DNA. The length of the gene open reading frame is 3,033 bp, predicting a protein of about 110 kDa. The sequence of this protein shares 10 blocks of homology with other eukaryotic ATPases, including the putative phosphorylation site characteristic of P-ATPases. Its hydropathy profile reveals 8-10 potential membrane-spanning regions. While the amino acid sequence of the T. brucei ATPase shows only 25% overall homology with its counterpart from the related kinetoplastid protozoan Leishmania donovani, 49% sequence conservation is found when compared with the calcium-ATPase from rabbit sarcoplasmic reticulum. This gene is present in only one copy, localized in the large chromosome fraction. It is transcribed at a similar level in procyclic and bloodstream forms, as a 4.3-kb mRNA. Run-on assays suggest continuous transcription of the gene and flanking sequences over at least 10 kb, by a RNA polymerase sensitive to alpha-amanitin. Transcription inhibition by UV irradiation suggests that the ATPase gene is more than 4 kb downstream from its promoter.
Revelard
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Department of Molecular Biology, University of Brussels, Rhode Saint Genèse, Belgium.
Pays
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D000546:Amanitins / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D000252:Calcium-Transporting ATPases / Q000235:genetics* / Q000378:metabolism; D015342:DNA Probes; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D015870:Gene Expression; D007893:Leishmania donovani / Q000201:enzymology / Q000235:genetics; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000201:enzymology; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic; D014346:Trypanosoma brucei brucei / Q000201:enzymology / Q000235:genetics*
D000546:Amanitins; D015342:DNA Probes; D012321:DNA-Directed RNA Polymerases; D000252:Calcium-Transporting ATPases