Bienz M
EMBO J. 1984 Nov;3(11):2477-83. doi: 10.1002/j.1460-2075.1984.tb02159.x.
Xenopus heat-shock genes are transiently heat-inducible in somatic cells, but they are also subject to a long-term developmental control in oogenesis and early embryogenesis. In order to understand whether different genes or different promoter elements are involved in the two types of control, several genomic clones coding for Xenopus heat-shock proteins, hsp 70 and hsp 30, were isolated, characterised and tested for expression in oocytes and COS cells. Three isolated hsp 70 genes are nearly identical in their promoter and mRNA leader sequences, indicating that there is only one type of hsp 70 gene. These promoters contain a consensus sequence element (CT-GAA--TTC-AG) upstream of the TATA-box, which is presumably required for their transient heat-inducibility. The two isolated hsp 30 genes show 5'-flanking sequences similar to each other, except that one of them shows a homology disruption precisely around the consensus sequence element. The same gene contains a frameshift mutation in the protein coding part and, since it cannot be expressed after introduction into oocytes or COS cells, it is probably a pseudogene. The other hsp 30 gene is strongly heat-inducible in injected oocytes or transfected COS cells. In contrast, the hsp 70 genes are strongly heat-inducible in COS cells, but their expression is highly efficient in injected oocytes at the normal temperature and is not increased during heat shock. This represents correct cell type-specific regulation of a cloned reintroduced gene, since the endogenous hsp 70 genes are constitutively activated during oogenesis, leading to the accumulation of stored hsp 70 mRNA in oocytes.
非洲爪蟾热休克基因在体细胞中可被热短暂诱导,但在卵子发生和早期胚胎发生过程中也受到长期的发育调控。为了了解两种调控方式是否涉及不同的基因或不同的启动子元件,我们分离、鉴定了几个编码非洲爪蟾热休克蛋白hsp 70和hsp 30的基因组克隆,并检测了它们在卵母细胞和COS细胞中的表达情况。分离出的三个hsp 70基因的启动子和mRNA前导序列几乎相同,这表明只有一种类型的hsp 70基因。这些启动子在TATA框上游含有一个共有序列元件(CT-GAA--TTC-AG),推测这是它们短暂热诱导所必需的。分离出的两个hsp 30基因的5'侧翼序列彼此相似,只是其中一个在共有序列元件周围恰好出现了同源性破坏。同一个基因在蛋白质编码部分存在移码突变,并且由于将其导入卵母细胞或COS细胞后无法表达,它可能是一个假基因。另一个hsp 30基因在注射的卵母细胞或转染的COS细胞中可被强烈热诱导。相比之下,hsp 70基因在COS细胞中可被强烈热诱导,但其在正常温度下注射的卵母细胞中表达效率很高,且在热休克期间不会增加。这代表了对克隆的重新导入基因的正确细胞类型特异性调控,因为内源性hsp 70基因在卵子发生过程中被组成性激活,导致卵母细胞中储存的hsp 70 mRNA积累。