Dimitriadis G J, Tata J R
Biochem J. 1982 Feb 15;202(2):491-7. doi: 10.1042/bj2020491.
The sensitivity to DNAase (deoxyribonuclease) I (which preferentially digests transcribed sequences) of vitellogenin and albumin genes in liver and erythrocytes of male Xenopus after primary and secondary induction of vitellogenesis by oestrogen was measured by hybridization to cDNA (complementary DNA) of the residual DNA after enzymic digestion of isolated nuclei. Vitellogenin sequences were rendered selectively more sensitive to limited DNAase-I digestion (15-20% of DNA rendered acid-soluble) during primary hormonal activation (5 days) of vitellogenin genes in liver, but not erythrocyte, nuclei. Hormone withdrawal (25 days after first injection) did not result in reversion to a pre-activation gene configuration, nor did secondary hormonal stimulation (5 days after second and 25 days after first injection) augment the sensitivity of the genes to digestion by the nuclease. Similar hormone treatment did not affect the sensitivity of the constitutively expressed albumin genes in liver nuclei, nor their insensitivity in erythrocyte nuclei. Under the same conditions, globin genes remained indigestible in liver nuclei. It is concluded that primary induction of vitellogenesis in male Xenopus liver is accompanied by relatively long-lasting (3-4 weeks) change in the configuration of vitellogenin genes in hepatic nuclei which is not reversed or further modified during short-term oestrogen withdrawal or upon secondary stimulation.
通过将分离细胞核经酶消化后的残余DNA与cDNA(互补DNA)杂交,测定了雌激素对非洲爪蟾雄性肝脏和红细胞中卵黄蛋白原基因及白蛋白基因进行初次和二次卵黄生成诱导后对DNA酶I(优先消化转录序列)的敏感性。在肝脏而非红细胞细胞核中,卵黄蛋白原基因在初次激素激活(5天)期间,卵黄蛋白原序列对有限的DNA酶I消化(15 - 20%的DNA变为酸溶性)选择性地变得更敏感。激素撤除(首次注射后25天)并未导致恢复到激活前的基因构型,二次激素刺激(第二次注射后5天和首次注射后25天)也未增强基因对核酸酶消化的敏感性。类似的激素处理不影响肝脏细胞核中组成型表达的白蛋白基因的敏感性,也不影响其在红细胞细胞核中的不敏感性。在相同条件下,珠蛋白基因在肝脏细胞核中仍不可消化。结论是,非洲爪蟾雄性肝脏中卵黄生成的初次诱导伴随着肝细胞核中卵黄蛋白原基因构型相对持久(3 - 4周)的变化,在短期雌激素撤除或二次刺激期间不会逆转或进一步改变。