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Nucleic Acids Research, 1984, Vol. 12, No. 24 9457-9472
© 1984

MOLECULAR BIOLOGY

{alpha}-Amanitin-insensitive transcription of variant surface glycoprotein genes provides further evidence for discontinuous transcription in trypanosomes

Jan M. Kooter and Piet Borst

Division of Molecular Biology, Antoni van Leeuwenhoek Huis, Netherlands Cancer Institute Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands

Received November 12, 1984. Accepted November 22, 1984.

Many, if not all, mRNAs in T.brucei start with the same sequence of 35 nucleotides, separately encoded in clustered so-called mini-exon repeats. From these mini-axon repeats a 141-nt precursor RNA with the 35-nt sequence at its 5'end is transcribed. Indirect evidence suggests that this RNA is linked in a second step to pre-mRNA transcripts. We have studied the sensitivity of RNA synthesis to {alpha}-amanitin in isolated trypanosome nuclei. Transcription of several protein coding genes is almost completely inhibited by a concentration of 5 µg {alpha}-amanitin per ml, whereas strong inhibition of mini-exon transcription is achieved with 200 µg {alpha}-amanitin per ml. In contrast, transcription of genes for variant surface glycoproteins (VSGs) is not inhibited by 1000 µg {alpha}-amanitin per ml, as is transcription of the genes for the major rRNAs. Since the mature VSG mRNAs start with the 35-nt sequence, our results provide additional evidence that the 35-nt sequence and the main part of VSG mRNA are produced from independent transcription units, these are transcribed by (partly) different RNA polymerases.


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