出版社：American Society for Biochemistry and Molecular Biology
摘要：To better characterize the increase in lipoprotein lipase (LPL) translation by hypothyroidism, adipocytes were prepared from control and hypothyroid rats. Whereas LPL synthesis was higher in hypothyroid adipocytes, with no change in mRNA levels, there was no increase in hormone-sensitive lipase (HSL) synthesis. To determine whether a transacting translation regulatory factor was present, a cytoplasmic fraction was prepared from control and hypothyroid adipocytes, and added to an in vitro translation system containing the hLPL mRNA. The hypothyroid cell fraction from adipose and heart yielded an increase in LPL translation, when compared to control extracts. Further experiments determined that the control adipocyte extract contained a translation-inhibitory factor that was 8-fold lower in activity in the hypothyroid extract. Using different LPL mRNA constructs in the in vitro translation reaction, the region that controlled translation was localized to nucleotides 1599 to 1638 (proximal 3' untranslated region (UTR)). To confirm the presence of a transacting factor, a sense RNA strand corresponding to this region was added to the in vitro translation reaction. This sense strand competed for the transacting factor in the control cell extract, yet had no effect on the hypothyroid cell extract. Thus, there is a translation repressor factor in the cytoplasm of rat adipocytes, and this factor is greatly reduced in activity in hypothyroid rat adipocytes. Because a similar mechanism of LPL regulation occurs in response to epinephrine, the absence of the translation repressor may be a mechanism for the loss of sensitivity of hypothyroid cells for catecholamines.
