Imboden M A, Laird P W, Affolter M, Seebeck T
Institute of General Microbiology, Bern, Switzerland.
Nucleic Acids Res. 1987 Sep 25;15(18):7357-68. doi: 10.1093/nar/15.18.7357.
The tubulin genes of T. brucei are clustered in a tightly packed array of alternating alpha- and beta-genes. The steady state mRNA contains one abundant mRNA species each for alpha- and beta-tubulin, both carrying the identical 35 nt mini-exon sequence at their 5'-ends. We have used in vitro run-on transcription assays to investigate the mechanism of tubulin gene transcription in T. brucei. Our results show that the regions between the individual tubulin genes are transcribed at the same rate as are the genes themselves. On the other hand, transcripts containing the intergenic regions could not be detected by Northern analysis or in vivo labelling experiments. We conclude that putative transcripts from the intergenic regions have a half-life of less than one minute. These results suggest that the tubulin gene cluster is transcribed as a single contiguous transcription unit yielding a primary transcript which is rapidly processed into individual mRNAs by the polyadenylation and mini-exon trans splicing machineries.
布氏锥虫的微管蛋白基因以紧密排列的α-基因和β-基因交替阵列的形式聚集在一起。稳态mRNA中α-和β-微管蛋白各有一个丰富的mRNA种类,两者在其5'-末端都携带相同的35个核苷酸的小外显子序列。我们利用体外连续转录分析来研究布氏锥虫微管蛋白基因的转录机制。我们的结果表明,各个微管蛋白基因之间的区域转录速率与基因本身相同。另一方面,通过Northern分析或体内标记实验无法检测到包含基因间区域的转录本。我们得出结论,基因间区域的推定转录本半衰期小于一分钟。这些结果表明,微管蛋白基因簇作为一个单一的连续转录单元进行转录,产生一个初级转录本,该转录本通过多聚腺苷酸化和小外显子反式剪接机制迅速加工成单个mRNA。