Bond U, Schlesinger M J
Mol Cell Biol. 1986 Dec;6(12):4602-10. doi: 10.1128/mcb.6.12.4602-4610.1986.
A chicken genomic library was screened to obtain genomic clones for ubiquitin genes. Two genes that differ in their genomic location and organization were identified. One gene, designated Ub I, contains four copies of the protein-coding sequence arranged in tandem, while the second gene, Ub II, contains three. The origin of the two major mRNAs that are induced after heat shock in chicken embryo fibroblasts was determined by generating DNA probes from the 5'-and 3'-noncoding regions of the two genes. Both mRNAs are transcribed from Ub I, the larger being the unspliced precursor of the smaller. A 674-base-pair intron was located within the 5'-noncoding region of Ub I. The second gene, Ub II, does not appear to code for an RNA species in normal or heat-shocked chicken embryo fibroblasts. The expression of ubiquitin mRNA during heat shock and recovery was examined. Addition of actinomycin D before heat shock completely abolished the response of ubiquitin mRNA to the stress. Analysis of the stability of the mRNA during recovery revealed that the mRNA accumulated during the heat shock is rapidly degraded with a half-life of approximately 1.5 h, suggesting a specialized but transient role for ubiquitin during heat shock.
通过筛选鸡基因组文库来获得泛素基因的基因组克隆。鉴定出两个在基因组位置和组织上不同的基因。一个基因命名为Ub I,包含四个串联排列的蛋白质编码序列拷贝,而第二个基因Ub II包含三个。通过从这两个基因的5'和3'非编码区生成DNA探针,确定了鸡胚成纤维细胞热休克后诱导产生的两种主要mRNA的来源。两种mRNA均由Ub I转录而来,较大的是较小的未剪接前体。在Ub I基因的5'非编码区内发现了一个674个碱基对的内含子。第二个基因Ub II在正常或热休克的鸡胚成纤维细胞中似乎不编码RNA种类。研究了热休克和恢复过程中泛素mRNA的表达。热休克前添加放线菌素D完全消除了泛素mRNA对应激的反应。对恢复过程中mRNA稳定性的分析表明,热休克期间积累的mRNA迅速降解,半衰期约为1.5小时,这表明泛素在热休克期间具有特殊但短暂的作用。