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首页> 美国卫生研究院文献>Molecular and Cellular Biology >Characterization of the human lipoprotein lipase (LPL) promoter: evidence of two cis-regulatory regions LP-alpha and LP-beta of importance for the differentiation-linked induction of the LPL gene during adipogenesis.
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Characterization of the human lipoprotein lipase (LPL) promoter: evidence of two cis-regulatory regions LP-alpha and LP-beta of importance for the differentiation-linked induction of the LPL gene during adipogenesis.

机译：人类脂蛋白脂肪酶（LPL）启动子的特征：两个顺式调节区域LP-α和LP-β的证据对脂肪形成过程中LPL基因的分化相关诱导很重要。

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摘要

When preadipocytes differentiate into adipocytes, several differentiation-linked genes are activated. Lipoprotein lipase (LPL) is one of the first genes induced during this process. To investigate early events in adipocyte development, we have focused on the transcriptional activation of the LPL gene. For this purpose, we have cloned and fused different parts of intragenic and flanking sequences with a chloramphenicol acetyltransferase reporter gene. Transient transfection experiments and DNase I hypersensitivity assays indicate that several positive as well as negative elements contribute to transcriptional regulation of the LPL gene. When reporter gene constructs were stably introduced into preadipocytes, we were able to monitor and compare the activation patterns of different promoter deletion mutants at selected time points representing the process of adipocyte development. We could delimit two cis-regulatory elements important for gradual activation of the LPL gene during adipocyte development in vitro. These elements, LP-alpha (-702 to -666) and LP-beta (-468 to -430), contain a striking similarity to a consensus sequence known to bind the transcription factors HNF-3 and fork head. Results of gel mobility shift assays and DNase I and exonuclease III in vitro protection assays indicate that factors with DNA-binding properties similar to those of the HNF-3/fork head family of transcription factors are present in adipocytes and interact with LP-alpha and LP-beta. We also demonstrate that LP-alpha and LP-beta were both capable of conferring a differentiation-linked expression pattern to a heterolog promoter, thus mimicking the expression of the endogenous LPL gene during adipocyte differentiation. These findings indicate that interactions with LP-alpha and LP-beta could be a part of a differentiation switch governing induction of the LPL gene during adipocyte differentiation.
机译：当前脂肪细胞分化为脂肪细胞时，几个分化相关基因被激活。脂蛋白脂肪酶（LPL）是在此过程中诱导的首批基因之一。为了调查脂肪细胞发育中的早期事件，我们集中于LPL基因的转录激活。为此，我们已将氯霉素乙酰基转移酶报道基因克隆并融合了基因内和侧翼序列的不同部分。瞬时转染实验和DNase I超敏性检测表明，几种阳性和阴性成分均对LPL基因的转录调控有贡献。当将报告基因构建体稳定地引入前脂肪细胞中时，我们能够在代表脂肪细胞发育过程的选定时间点监测和比较不同启动子缺失突变体的激活模式。我们可以界定两个顺式调控元件，这些元件在体外脂肪细胞发育过程中对LPL基因的逐步激活很重要。这些元件LP-alpha（-702至-666）和LP-beta（-468至-430）与已知的结合转录因子HNF-3和叉头的共有序列具有惊人的相似性。凝胶迁移率移动分析以及DNase I和核酸外切酶III体外保护分析的结果表明，脂肪细胞中存在与HNF-3 /叉头转录因子家族相似的DNA结合特性因子，并与LP-alpha和LP-beta。我们还证明了LP-α和LP-β都能够将分化相关的表达模式赋予异源启动子，从而模拟脂肪细胞分化过程中内源LPL基因的表达。这些发现表明与LP-α和LP-β的相互作用可能是在脂肪细胞分化过程中控制LPL基因诱导的分化开关的一部分。

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