Pierrat B, Lacroute F, Losson R
Laboratoire of Génétique Moléculaire des Eucaryotes du CNRS, Strasbourg, France.
Gene. 1993 Sep 6;131(1):43-51. doi: 10.1016/0378-1119(93)90667-r.
In Saccharomyces cerevisiae, the mRNA encoded by the PPR1 gene is very unstable (t1/2 = 1 min), whereas the mRNA encoded by the URA3 gene is relatively stable (t1/2 = 10 min). To identify cis-acting sequences that dictate mRNA decay rates in yeast, we have constructed PPR1/URA3 gene fusions and measured the half-lives of the resulting chimeric transcripts. The mRNA containing the URA3 coding region fused to the untranslated regions (UTR) of PPR1 decayed at a rate similar to the native PPR1 mRNA, suggesting that the instability of the PPR1 mRNA is not linked to its coding sequence. When the 5'-UTR of PPR1 was replaced by the 5'-UTR of URA3, the chimeric transcript was strongly stabilized, indicating that the 5'-UTR of PPR1 is required for the rapid decay of its mRNA. Fusion of this PPR1 5'-UTR to the URA3 coding region was sufficient to destabilize the chimeric mRNA. We conclude that the PPR1 5'-UTR contains sequence(s) that can promote rapid mRNA decay in yeast.
在酿酒酵母中,PPR1基因编码的mRNA非常不稳定(半衰期t1/2 = 1分钟),而URA3基因编码的mRNA相对稳定(半衰期t1/2 = 10分钟)。为了鉴定决定酵母中mRNA降解速率的顺式作用序列,我们构建了PPR1/URA3基因融合体,并测定了所得嵌合转录本的半衰期。含有与PPR1非翻译区(UTR)融合的URA3编码区的mRNA以与天然PPR1 mRNA相似的速率降解,这表明PPR1 mRNA的不稳定性与其编码序列无关。当PPR1的5'-UTR被URA3的5'-UTR取代时,嵌合转录本得到强烈稳定,表明PPR1的5'-UTR是其mRNA快速降解所必需的。将该PPR1 5'-UTR与URA3编码区融合足以使嵌合mRNA不稳定。我们得出结论,PPR1的5'-UTR包含能够促进酵母中mRNA快速降解的序列。