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首页> 外文期刊>The FEBS journal >Nuclear factor kappa B and tumor necrosis factor-alpha modulation of transcription of the mouse testis- and pre-implantation development-specific Rnf33/Trim60 gene
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Nuclear factor kappa B and tumor necrosis factor-alpha modulation of transcription of the mouse testis- and pre-implantation development-specific Rnf33/Trim60 gene

机译：小鼠睾丸和植入前发育特异性Rnf33 / Trim60基因转录的核因子κB和肿瘤坏死因子-α调制

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摘要

We have previously reported a mouse Rnf33/Trim60 gene that is temporally expressed in the pre-implantation embryo. The Rnf33 structural gene is composed of a short noncoding exon 1 and an intronless coding exon 2. In the present work, Rnf33 was shown to be expressed in the mouse testis and in the testicular cell lines TM3 and TM4. To elucidate Rnf33 transcriptional modulation, a 2.5-kb Rnf33 sequence, inclusive of the upstream regulatory region, exon 1 and the associated intronic sequence, was dissected in transient transfection and luciferase assays. An initiator and an atypical TATA-box were shown to act as the core promoter elements of the gene. Deletion and mutagenesis of the 2.5-kb sequence in luciferase constructs further demonstrated that an intronic and palindromic kappa B (kappa B) sequence was an important cis element targeted by the nuclear factor-kappa B (NF-kappa B) subunits p65/RELA and p50/NF kappa B1, and also through modulation by tumor necrosis factor alpha. Transcriptional up-regulation of Rnf33 by NF-kappa B and tumor necrosis factor-alpha was directly demonstrated in TM3 and TM4 cells by real-time PCR quantification of the Rnf33 mRNA levels. Small interfering RNA knockdown of p65 and p50 confirmed Rnf33 down-regulation by p65/p50. Spermatogenesis is regulated by a wide range of stimuli, including NF-kappa B, which, in turn, is regulated by other signals. Hence, demonstration of NF-kappa B-regulated Rnf33 expression in testicular cells, particularly in Sertoli cells, implicates functional involvement of the putative RNF33 protein in spermatogenesis through association of the RNF33 protein with the microtubule via interaction with kinesin motor proteins, as previously demonstrated [Huang et al., submitted].
机译：我们以前已经报道过小鼠Rnf33 / Trim60基因在植入前的胚胎中暂时表达。 Rnf33结构基因由短的非编码外显子1和无内含子编码外显子2组成。在本工作中，Rnf33已显示在小鼠睾丸以及睾丸细胞系TM3和TM4中表达。为了阐明Rnf33的转录调控，在瞬时转染和萤光素酶测定中解剖了一个2.5-kb的Rnf33序列，包括上游调节区，外显子1和相关的内含子序列。已显示启动子和非典型的TATA盒可作为基因的核心启动子元件。荧光素酶构建物中2.5kb序列的缺失和诱变进一步证明，内含子和回文Kappa B（kappa B）序列是核因子-κB（NF-kappa B）亚基p65 / RELA和p65 / RELA靶向的重要顺式元件。 p50 / NF kappa B1，也可以通过肿瘤坏死因子α调节。通过实时PCR定量Rnf33 mRNA水平，在TM3和TM4细胞中直接证明了NF-κB和肿瘤坏死因子-α对Rnf33的转录上调。 p65和p50的小干扰RNA敲低证实了p65 / p50对Rnf33的下调。精子发生受到包括NF-κB在内的各种刺激的调节，而NF-κB又受其他信号的调节。因此，如前所述，在睾丸细胞中，特别是在睾丸支持细胞中，NF-κB调节Rnf33的表达表明，假定的RNF33蛋白通过与驱动蛋白运动蛋白相互作用，通过RNF33蛋白与微管的结合而参与精子发生。 [Huang等，已提交]。

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