Sparks K A, Dieckmann C L
Department of Biochemistry and Department Molecular and Cellular Biology, Life Sciences South Room 454, University of Arizona, Tucson, AZ 85721, USA.
Nucleic Acids Res. 1998 Oct 15;26(20):4676-87. doi: 10.1093/nar/26.20.4676.
Several yeast genes produce multiple transcripts with different 3'-ends. Of these, four genes are known to produce truncated transcripts that end within the coding sequence of longer transcripts: CBP1 , AEP2 / ATP13 , RNA14 and SIR1 . It has been shown that the level of the truncated CBP1 transcript increases during the switch to respiratory growth while that of the full-length transcript decreases. To determine whether this phenomenon is unique to CBP1 , northern analysis was used to determine whether the levels of other truncated transcripts are regulated similarly by carbon source. The levels of the shortest transcripts of AEP2 / ATP13 and RNA14 increased during respiration while the shortest SIR1 transcript remained constant. However, two longer SIR1 transcripts were regulated reciprocally by carbon source. Mapping the 3'-ends of each transcript by sequencing partial cDNA clones revealed multiple 3'-ends for each transcript. Examination of the sequences surrounding the 3'-ends of the induced transcripts failed to identify a consensus sequence but did reveal weak putative 3'-end formation signals in all of the transcripts. Similarly, no consensus sequence was found when the sequences surrounding the 3'-ends of the longest transcripts were compared, but again weak putative 3'-end formation signals were identified. These data are suggestive of carbon source regulation of alternative poly(A) site choice in yeast.
几个酵母基因可产生具有不同3'末端的多种转录本。其中,已知有四个基因会产生在较长转录本编码序列内终止的截短转录本:CBP1、AEP2 / ATP13、RNA14和SIR1。研究表明,在转换为呼吸生长过程中,截短的CBP1转录本水平升高,而全长转录本水平降低。为了确定这种现象是否为CBP1所特有,采用Northern印迹分析来确定其他截短转录本的水平是否受碳源的类似调控。在呼吸过程中,AEP2 / ATP13和RNA14最短转录本的水平升高,而最短的SIR1转录本保持恒定。然而,两个较长的SIR1转录本受碳源的反向调控。通过对部分cDNA克隆进行测序来定位每个转录本的3'末端,结果显示每个转录本都有多个3'末端。对诱导转录本3'末端周围的序列进行检查,未能鉴定出共有序列,但确实在所有转录本中发现了微弱的假定3'末端形成信号。同样,比较最长转录本3'末端周围的序列时也未发现共有序列,但再次鉴定出了微弱的假定3'末端形成信号。这些数据表明酵母中可变聚腺苷酸化位点的选择受碳源调控。