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Expression of the Thimet Oligopeptidase Gene is Regulated by Positively and Negatively Acting Elements

机译：Thimet寡肽酶基因的表达受正负作用元件的调节。

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摘要

Thimet oligopeptidase (TOP) is a thiol-dependent metallopeptidase, which can cleave and thereby modulate the activity of many neuropeptides. The enzyme is active in many endocrine tissues, including testis, brain, and pituitary. In rat, the richest source of TOP is the testes, with a specific activity fivefold that of brain. The mechanism whereby rat TOP expression is regulated at the transcriptional level has been examined by reporter gene assay and electromobility shift assays after isolation of 1020 bp of upstream sequence. Computer analysis predicts a number of potential transcription factor-binding sites, which were examined by deletion analysis and DNA-binding studies. The promoter or its deletion fragments were fused to luciferase reporter gene vectors and introduced into GH3 pituitary, COS-1 kidney, MAT-Lu prostate, and GC-2spd(ts) spermatid cells. Two regions of the promoter have been identified: a positively acting region (-901/-219) and a strong negatively acting region (-219/-102). Concomitantly, potential transcription factors interacting with the cis-acting elements of the promoter were studied by gel electromobility shift assays. This work has identified a number of transcription factor-binding sites. However, no differences in the binding behavior in the various cell lines was observed.
机译：Thimet寡肽酶（TOP）是一种硫醇依赖性金属肽酶，可以裂解并调节许多神经肽的活性。该酶在许多内分泌组织（包括睾丸，大脑和垂体）中都有活性。在大鼠中，TOP的最丰富来源是睾丸，其比活性是大脑的五倍。分离出上游序列的1020 bp后，通过报道基因基因检测和电动迁移检测分析了大鼠TOP表达在转录水平上调控的机制。计算机分析预测了许多潜在的转录因子结合位点，这些位点通过缺失分析和DNA结合研究进行了检查。将启动子或其缺失片段与萤光素酶报道基因载体融合，并引入到GH3垂体，COS-1肾脏，MAT-Lu前列腺和GC-2spd（ts）精子细胞中。已经鉴定出启动子的两个区域：正作用区域（-901 / -219）和强负作用区域（-219 / -102）。同时，通过凝胶电迁移率测定研究了与启动子的顺式作用元件相互作用的潜在转录因子。这项工作已经确定了许多转录因子结合位点。然而，在各种细胞系中未观察到结合行为的差异。

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