Puzianowska-Kuznicka M, Shi Y B
Laboratory of Molecular Embryology, NICHD, National Institutes of Health, Bethesda, Maryland 20892-5430, USA.
J Biol Chem. 1996 Mar 15;271(11):6273-82. doi: 10.1074/jbc.271.11.6273.
Nuclear factor I (NFI) family members are transcription factors that are believed to also participate in DNA replication. We have cloned two Xenopus laevis NFIs that are up-regulated by thyroid hormone. They are 84-95% identical to their counterparts in birds and mammals. In contrast, the two Xenopus NFIs are much less homologous to each other, sharing only 58% homology, which largely resides in the DNA binding domain at the amino terminus. However, both NFIs can bind to a consensus NFI binding site and activate the transcription of a promoter bearing the site. Northern blot reveals that both NFI genes are regulated in tissue- and developmental stage-dependent manners. They are first activated, independently of thyroid hormone, to low levels at stages 23/24, around the onset of larval organogenesis. After stage 54, their mRNA levels are dramatically upregulated by endogenous thyroid hormone, and high levels of their expression correlate with organ-specific metamorphosis. Furthermore, gel mobility shift assay indicates that the NFI proteins are present in different organs and that their levels are regulated similarly to the mRNA levels. These results strongly suggest that NFIs play important roles during postembryonic organ development, in contrast to the general belief that NFIs are ubiquitous factors.
核因子I(NFI)家族成员是转录因子,据信也参与DNA复制。我们克隆了两种受甲状腺激素上调的非洲爪蟾NFI。它们与鸟类和哺乳动物中的对应物有84 - 95%的同一性。相比之下,这两种非洲爪蟾NFI彼此之间的同源性要低得多,仅共享58%的同源性,且主要存在于氨基末端的DNA结合结构域中。然而,两种NFI都能结合到一致的NFI结合位点并激活带有该位点的启动子的转录。Northern印迹显示,两个NFI基因都以组织和发育阶段依赖性方式受到调控。它们在第23/24阶段,即幼虫器官发生开始时,首先独立于甲状腺激素被激活至低水平。在第54阶段之后,它们的mRNA水平被内源性甲状腺激素显著上调,且其高表达水平与器官特异性变态相关。此外,凝胶迁移率变动分析表明,NFI蛋白存在于不同器官中,且其水平与mRNA水平的调控方式相似。这些结果有力地表明,与普遍认为NFI是普遍存在的因子的观点相反,NFI在胚胎后器官发育过程中发挥重要作用。