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Nucleic Acids Research, 1992, Vol. 20, No. 11 2725-2734
© 1992

MOLECULAR BIOLOGY

A ribosomal RNA gene promoter at the telomere of a mini-chromosome in Trypanosoma brucei

Joost C.B.M. Zomerdijk, Rudo Kieft and Piet Borst*

Division of Molecular Biology, The Netherlands Cancer Institute Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands

*To whom correspondence should be addressed

Received March 16, 1992. Accepted April 28, 1992.

The parasitic protozoan Trypanosoma brucei has some hundred mini-chromosomes of 50–150 kb, which mainly consist of telomeric repeats, sub-telomeric repeats and internal 177-bp repeats. Their primary function seems to be to expand the repertoire of non-transcribed sub-telomeric variant surface glycoprotein (VSG) genes. Here we report that two of the smaller mini-chromosomes (55 and 60 kb) contain sequences homologous to the ribosomal RNA gene promoter region. We have targeted by homologous recombination the neomycin phosphotransferase (neo1) gene behind the promoter on the 55 kb chromosome and show that this promoter mediates the efficient synthesis of properly trans-spliced and polyadenylated neo mRNA. The resulting high resistance to G418 (a neo analogue) is stable in the absence of drug showing that mitotic segregation of this mini-chromosome is precise. Downstream of the transcription start the wild-type version of the ribosomal promoter is flanked by telomeric repeats. The absence of the sub-telomeric repeats found in other T.brucel chromosome ends suggests that the rDNA-telomere junction has been formed by de novo addition of telomeric repeats to a broken chromosome end (healing). Our results provide a plausible explanation for the {alpha}-amanltin-resistant transcription of telomeric repeats in T.brucei reported by Rudenko and Van der Ploeg1 and they show that trypanosomes can efficiently use RNA polymerase I for the expression of sub-telomeric genes, supporting the notion that the {alpha}-amanitin-resistant transcription of sub-telomeric VSG genes may also be catalyzed by this enzyme.


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