Cloning and characterization of human cathepsin L promoter
Cloning and characterization of human cathepsin L promoter
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DOI：
10.1016/s0378-1119(01)00650-3
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发表时间：
2001-09-05
期刊：
影响因子：
3.5
通讯作者：
Chauhan, SS
中科院分区：
文献类型：
--
作者：
Bakhshi, R;Goel, A;Chauhan, SS
组织蛋白酶L是一种溶酶体半胱氨酸蛋白酶，由恶性细胞过度表达和分泌。它在降解胶原蛋白、弹性蛋白、层粘连蛋白和基底膜的其他组分方面非常有效，因此，与肿瘤侵袭和转移有关。克隆人组织蛋白酶L（hCATL）基因的结构部分以阐明其基因组组织（Chauhan et al.，J.Biol.Chem.218（1993）1039）。本研究利用PCR技术从连接接头的胎盘基因组文库中扩增出一个1.90kb的DNA片段，该片段包含hCATL的5'上游区1825 bp和第一外显子75个碱基。通过荧光素酶报告基因测定已经证明该片段表现出启动子活性。该片段的序列分析揭示了大约29个不同的推定的转录因子结合位点的存在。其中AP-4、加塔-1、Lmo 2、CEBPB、MZF-1、NFAT等几种在该区域出现不止一次。然而，在外显子1上游1.0 kb内发现了共有CAAT盒，但没有发现共有TATA盒。使用胎盘总RNA，将hCATL的转录起始位点定位于ATG密码子上游289个碱基的单个腺嘌呤残基。(C)2001 Elsevier Science B. V.保留所有权利。
Cathepsin L is a lysosomal cysteine protease, which is over-expressed and secreted by malignant cells. It is very potent in degrading collagen, elastin, laminin and other components of the basement membrane and, therefore, has been implicated in tumor invasion and metastasis. The structural portion of the human cathepsin L (hCATL) gene was cloned to elucidate its genomic organization (Chauhan et al., J. Biol. Chem. 218 (1993) 1039). In the present study, a 1.90 kb DNA fragment, containing 1825 bp of the 5' upstream region of hCATL and 75 bases of the first exon of the hCATL, was amplified by PCR from an adaptor ligated placental genomic library. This fragment has been demonstrated to exhibit promoter activity by luciferase reporter assays. Sequence analysis of this fragment revealed the presence of approximately 29 different putative transcription factor binding sites. Several of them like AP-4, GATA-1, Lmo2, CEBPB, MZF-1, NFAT, etc. were present more than once in this region. However, a consensus CAAT box but no consensus TATA box was found within the 1.0 kb upstream of exon 1. The transcription initiation site of hCATL, using placental total RNA, was mapped to a single adenine residue 289 bases upstream of the ATG codon. (C) 2001 Elsevier Science B.V. All rights reserved.