The human iodotyrosine dehalogenase 1 (DEHAL1) gene is composed of six exons. Two isoforms (DEHAL1 and DEHAL1B) have been published in GenBank, both of which have a nitroreductase domain and arise from differential splicing in exon 5. We recently showed that the DEHAL1 isoform is a transmembrane protein that efficiently catalyzes the NADPH-dependent deiodination of mono (L-MIT) and diiodotyrosine (L-DIT) in human embryonic kidney-293 (HEK293) cells. In the present study, we establish the existence of a new transcript, DEHAL1C, in the human thyroid with a terminal exon that lacks in the DEHAL1 transcript. This exon is the complete exon 5, which is spliced in the DEHAL1B mRNA variant. These two variants encode proteins with differing C-terminal domains. Using quantitative reverse transcription polymerase chain reaction, we found that the expression of the mRNA of DEHAL1C and DEHAL1B was lower than that of DEHAL1 mRNA in the thyroid. We also observed that human DEHAL1B and DEHAL1C proteins are rapidly degraded in stably transfected HEK293 cells, unlike the DEHAL1 protein, and that exposure to the proteasome inhibitor MG132 resulted in accumulation of these proteins that was markedly time- and concentration-dependent. These findings show that the cytoplasmic tail could play a role in the stability of the protein.

译文

人类碘酪氨酸脱卤酶1 (DEHAL1) 基因由六个外显子组成。GenBank上已发布了两种同工型 (DEHAL1和DEHAL1B),它们都具有硝基还原酶结构域,并且是由外显子5中的差异剪接引起的。我们最近发现DEHAL1亚型是一种跨膜蛋白,可有效催化人胚肾293 (HEK293) 细胞中mono (L-MIT) 和二碘酪氨酸 (L-DIT) 的NADPH依赖性脱碘作用。在本研究中,我们确定了人类甲状腺中存在一个新的转录本DEHAL1C,其末端外显子在DEHAL1转录本中缺乏。该外显子是完整的外显子5,在DEHAL1B mRNA变体中剪接。这两个变体编码具有不同C末端结构域的蛋白质。使用定量逆转录聚合酶链反应,我们发现DEHAL1C和DEHAL1B的mRNA在甲状腺中的表达低于DEHAL1 mRNA的表达。我们还观察到,与DEHAL1蛋白不同,人DEHAL1B和DEHAL1C蛋白在稳定转染的HEK293细胞中迅速降解,并且暴露于蛋白酶体抑制剂MG132导致这些蛋白的积累明显依赖于时间和浓度。这些发现表明,细胞质尾可能在蛋白质的稳定性中起作用。

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