Sun C W, Callis J
Section of Molecular and Cellular Biology, University of California, Davis 95616, USA.
Plant J. 1997 May;11(5):1017-27. doi: 10.1046/j.1365-313x.1997.11051017.x.
The highly conserved protein ubiquitin is encoded by five polyubiquitin genes in Arabidopsis thaliana ecotype Columbia that have been divided into two subtypes, the UBQ3/UBQ4 subtype and the UBQ10/UBQ11/UBQ14 subtype. Northern analysis using gene-specific oligonucleotides as hybridization probes and enzyme activity measurements from transgenic plants expressing beta-glucuronidase (GUS) under the control of individual polyubiquitin 5' flanking regions were used to determine the development and environmental regulation of polyubiquitin transcription and mRNA accumulation. Polyubiquitin mRNA levels within and between subtypes were independently modulated. UBQ3 mRNA levels were three-fold higher than UBQ4 mRNA levels in vegetative organs, but only two-thirds of the UBQ4 mRNA levels in flowers. UBQ3 mRNA was modulated by dark/light treatments, while mRNAs from UBQ and all members of the other subtype were unaffected. Similarly, within the UBQ10/UBQ11/UBQ14 subtype, UBQ11/UBQ14 mRNAs were modulated differently in seedlings after a two-hour heat-shock treatment. Among all the polyubiquitin genes, UBQ10 mRNA level was the most constant in all organs and environmental conditions examined. Transgenic plants transformed with a UBQ10 5' flanking region::GUS gene contained higher levels of GUS activity than transgenic plants expressing GUS under the control of UBQ3 5' flanking regions. In conclusion, the relative abundance of different Arabidopsis polyubiquitin mRNAs, even those produced from highly similar genes within a subtype, appears to be modulated independently in response to developmental and environmental cues.
高度保守的蛋白质泛素由拟南芥生态型哥伦比亚的五个多聚泛素基因编码,这些基因已被分为两个亚型,即UBQ3/UBQ4亚型和UBQ10/UBQ11/UBQ14亚型。使用基因特异性寡核苷酸作为杂交探针的Northern分析以及在各个多聚泛素5'侧翼区域控制下表达β-葡萄糖醛酸酶(GUS)的转基因植物的酶活性测量,用于确定多聚泛素转录和mRNA积累的发育和环境调控。亚型内部和之间的多聚泛素mRNA水平受到独立调节。在营养器官中,UBQ3 mRNA水平比UBQ4 mRNA水平高两倍,但在花中仅为UBQ4 mRNA水平的三分之二。UBQ3 mRNA受暗/光处理调节,而来自UBQ和其他亚型所有成员的mRNA不受影响。同样,在UBQ10/UBQ11/UBQ14亚型中,经过两小时热休克处理后,幼苗中的UBQ11/UBQ14 mRNA受到不同调节。在所有多聚泛素基因中,UBQ10 mRNA水平在所检查的所有器官和环境条件下最为稳定。用UBQ10 5'侧翼区域::GUS基因转化的转基因植物比在UBQ3 5'侧翼区域控制下表达GUS的转基因植物含有更高水平的GUS活性。总之,不同拟南芥多聚泛素mRNA的相对丰度,即使是那些由亚型内高度相似的基因产生的mRNA,似乎也会根据发育和环境线索进行独立调节。