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Expression of ferrochelatase mRNA in erythroid and non-erythroid cells.

机译:铁螯合酶mRNA在红系和非红系细胞中的表达。

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摘要

Ferrochelatase, which catalyses the last step in haem biosynthesis, i.e. the insertion of Fe(II) into protophorphyrin IX, is present in all cells, but is particularly abundant in erythroid cells during haemoglobinization. Using mouse ferrochelatase cDNA as a probe two ferrochelatase transcripts, having lengths of 2.9 kb and 2.2 kb, were found in extracts of mouse liver, kidney, brain, muscle and spleen, the 2.9 kb transcript being more abundant in the non-erythroid tissues and the 2.2 kb transcript more predominant in spleen. In mouse erythroleukemia cells the 2.9 kb ferrochelatase transcript is also more abundant; however, following induction of erythroid differentiation by dimethyl sulphoxide there is a preferential increase in the 2.2 kb transcript, which eventually predominates. With mouse reticulocytes, the purest immature erythroid cell population available, over 90% of the total ferrochelatase mRNA is present as the 2.2 kb transcript. Since there is probably only one mouse ferrochelatase gene, the occurrence of two ferrochelatase transcripts could arise from the use of two putative polyadenylation signals in the 3' region of ferrochelatase DNA. This possibility was explored by using a 389 bp DNA fragment produced by PCR with synthetic oligoprimers having sequence similarity with a region between the polyadenylation sites. This fragment hybridized only to the 2.9 kb ferrochelatase transcript, indicating that the two transcripts differ at their 3' ends and suggesting that the 2.2 kb transcript results from the utilization of the upstream polyadenylation signal. The preferential utilization of the upstream polyadenylation signal may be an erythroid-specific characteristic of ferrochelatase gene expression.
机译:铁螯合酶可催化血红素生物合成的最后一步,即将Fe(II)插入原卟啉IX中,存在于所有细胞中,但在血红蛋白化过程中在类红细胞中特别丰富。用小鼠亚铁螯合酶cDNA作为探针,在小鼠肝,肾,脑,肌肉和脾脏的提取物中发现了两种长度分别为2.9 kb和2.2 kb的亚铁螯合酶转录物,在非红系组织中该2.9 kb转录物更为丰富。 2.2 kb的转录本在脾脏中更占优势。在小鼠红白血病细胞中,2.9 kb的铁螯合酶转录物也更为丰富。但是,在通过二甲基亚砜诱导类红细胞分化之后,2.2 kb转录本会优先增加,最终占主导地位。对于小鼠网织红细胞,可获得的最纯的未成熟红系细胞群体,总铁螯合酶mRNA的90%以上以2.2 kb的转录本存在。由于可能只有一个小鼠亚铁螯合酶基因,因此在铁螯合酶DNA 3'区域使用两个推定的聚腺苷酸化信号可能会导致出现两个铁螯合酶转录物。通过使用PCR产生的389 bp DNA片段和合成的寡引物来探索这种可能性,该引物与聚腺苷酸化位点之间的区域具有序列相似性。该片段仅与2.9 kb的铁螯合酶转录物杂交,表明这两个转录物在其3'末端不同,这表明2.2 kb的转录物是利用上游多腺苷酸化信号产生的。上游聚腺苷酸化信号的优先利用可能是铁螯合酶基因表达的红系特异性特征。

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