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首页> 外文期刊>Cellular Physiology and Biochemistry >Regulation of Pokemon 1 activity by sumoylation
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Regulation of Pokemon 1 activity by sumoylation

机译：通过磺酰化调节口袋妖怪1的活动

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Pokemon 1 is a proto-oncogenic transcriptional regulator that contains a POZ domain at the N-terminus and four Kruppel-like zinc fingers at the C-terminus. Pokemon 1 plays an important role in adipogenesis, osteogenesis, oncogenesis, and transcription of NF-kB responsive genes. Recent reports have shown that biological activities of transcription factors are regulated by sumolylation. We investigated whether Pokemon 1 is post-translationally modified by sumoylation and whether the modification affects Pokemon 1's transcriptional properties. We found that Pokemon 1 is sumoylated in vitro and in vivo. Upon careful analysis of the amino acid sequence of Pokemon 1, we found ten potential sumoylation sites located at lysines 61, 354, 371, 379, 383, 396, 486, 487, 536 and 539. We mutated each of these amino acids into arginine and tested whether the mutation could affect the transcriptional properties of Pokemon 1 on the Pokemon 1 responsive genes, such as ADH5/FDH and pG5-FRE-Luc. Wild-type Pokemon 1 potently represses transcription of ADH5/FDH. Most of the mutants, however, were weaker transcription repressors and repressed transcription 1.3-3.3 fold less effective. Although potential sumoylation sites were located close to the DNA binding domain or the nuclear localization sequence, the mutations did not alter nuclear localization or DNA binding activity. In addition, on the pG5-FRE-Luc test promoter construct, ectopic SUMO-1 repressed transcription in the presence of Pokemon 1. The sumoylation target lysine residue at amino acid 61, which is located in the middle of the POZ-domain, is important because K61R mutation resulted in a much weaker molecular interaction with corepressors. Our data suggest that Pokemon 1's activity as a transcription factor may involve sumoylation, and that sumoylation might be important in the regulation of transcription by Pokemon 1.
机译：宠物小精灵1是原癌基因的转录调节因子，在N端包含一个POZ域，在C端包含四个Kruppel状锌指。宠物小精灵1在脂肪生成，成骨，肿瘤发生和NF-kB响应基因的转录中起重要作用。最近的报道表明，转录因子的生物活性受sumolylation调节。我们调查了口袋妖怪1是否通过sumoylation进行翻译后修饰，以及该修饰是否影响口袋妖怪1的转录特性。我们发现口袋妖怪1在体外和体内被磺酰化。通过仔细分析口袋妖怪1的氨基酸序列，我们发现位于赖氨酸61、354、371、379、383、396、486、487、536和539的10个潜在的磺酰化位点。我们将这些氨基酸各自突变为精氨酸。并测试了该突变是否会影响Pokemon 1响应基因（如ADH5 / FDH和pG5-FRE-Luc）上Pokemon 1的转录特性。野生型口袋妖怪1有效抑制ADH5 / FDH的转录。然而，大多数突变体是较弱的转录阻遏物，而被抑制的转录无效1.3-3.3倍。尽管潜在的磺酰化位点位于DNA结合域或核定位序列附近，但突变不会改变核定位或DNA结合活性。此外，在pG5-FRE-Luc测试启动子构建体上，异位SUMO-1在口袋妖怪1的存在下抑制了转录。位于POZ结构域中间的第61位氨基酸的sumoylation目标赖氨酸残基为：之所以重要，是因为K61R突变导致与共加压因子的分子相互作用弱得多。我们的数据表明，口袋妖怪1作为转录因子的活性可能涉及sumoylation，而sumoylation可能在口袋妖怪1的转录调控中很重要。

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