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首页> 外文期刊>Osteoarthritis and cartilage >Identification and differential expression of human collagenase-3 mRNA species derived from internal deletion, alternative splicing, and different polyadenylation and transcription initiation sites.
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Identification and differential expression of human collagenase-3 mRNA species derived from internal deletion, alternative splicing, and different polyadenylation and transcription initiation sites.

机译：鉴定和差异表达的人类胶原酶3 mRNA物种源自内部缺失，替代剪接以及不同的聚腺苷酸化和转录起始位点。

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OBJECTIVE: Collagenase-3 is a metalloprotease that plays a role in tissue remodeling and pathological processes including arthritis. The human gene is transcribed into major (3.0 and 2.5 kb) and minor (2.2/2.0 kb) transcripts, as seen in Northern blot assays. We investigated the possibility that other transcripts, not detectable by Northern blot, were synthesized as either coding or regulatory RNAs that would modulate collagenase-3 expression and function/activity. DESIGN: We screened a cDNA library and total RNA from human chondrocytes by plaque hybridization and RT-PCR, and expressed the transcripts in a cellular environment. The levels of expression of each transcript in normal and osteoarthritic joint cells and cartilage were monitored by RT-PCR. RESULTS: We identified five different collagenase-3 RNA species derived from alternative polyadenylation sites (COL3-APS), internal deletion (COL3-DEL), alternative splicing (COL3-9B/COL3-9B-2), and different transcription initiation site (COL3-ATS and COL3-ATS-INT). Each transcript could be translated in a cellular environment. Interestingly, the proteins translated from the COL3-DEL and COL3-9B-2 transcripts had a modified hemopexin-like domain, suggesting altered collagenolytic activities. The transcript types COL3-APS, COL3-9B-2, and COL3-ATS were up-regulated in the osteoarthritic samples and expressed in the chondrosarcoma cell line SW1353. CONCLUSION: Our data show that the human collagenase-3 gene is subjected to different levels of regulation and constitutes a more complex system than was originally thought.
机译：目的：胶原酶3是一种金属蛋白酶，在组织重塑和包括关节炎在内的病理过程中起作用。如Northern blot分析所示，人类基因被转录成主要（3.0和2.5 kb）和次要（2.2 / 2.0 kb）转录本。我们调查了通过RNA印迹无法检测到的其他转录本被合成为编码或调节RNA的可能性，这些转录本可以调节胶原酶3的表达和功能/活性。设计：我们通过噬菌斑杂交和RT-PCR筛选了人类软骨细胞的cDNA文库和总RNA，并在细胞环境中表达了转录本。通过RT-PCR监测正常和骨关节炎关节细胞和软骨中每个转录物的表达水平。结果：我们鉴定了五个不同的胶原酶3 RNA物种，分别来自替代的聚腺苷酸化位点（COL3-APS），内部缺失（COL3-DEL），替代剪接（COL3-9B / COL3-9B-2）和不同的转录起始位点（ （COL3-ATS和COL3-ATS-INT）。每个成绩单可以在细胞环境中翻译。有趣的是，从COL3-DEL和COL3-9B-2转录本翻译的蛋白质具有修饰的血红素样结构域，表明胶原蛋白水解活性发生了变化。在骨关节炎样品中，转录类型COL3-APS，COL3-9B-2和COL3-ATS上调并在软骨肉瘤细胞系SW1353中表达。结论：我们的数据表明，人类胶原酶3基因受到不同程度的调节，并且比原先所认为的构成了更为复杂的系统。

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