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Specific endonucleolytic cleavages of mouse albumin mRNA and their modulation during liver development.

机译:小鼠白蛋白mRNA的特异性内切核酸裂解及其在肝脏发育过程中的调控。

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摘要

We have detected specific endonucleolytic cleavages of mouse albumin mRNA by S1 nuclease protection analysis of total RNA from fetal mouse liver using a cDNA probe spanning the middle, coding region of albumin mRNA. With the use of probe labeled at its 5' end, three prominent cleavages were detected which were confirmed and their endonucleolytic nature was established by further analysis using 3' end-labeled probe. The latter probe also revealed one more cleavage which was not detected with the 5' end-labeled probe. These cleavages mapped to positions on the mRNA which included a unique sequence motif CCAN1-3CUGN0-1UGAU. Degradation intermediates corresponding to these cleavages were consistently observed, specifically in fetal liver but not in normal or regenerating adult liver and appeared to have originated in vivo. Their levels decreased progressively from 18th day of gestation and became undetectable by 20 days after birth. No detectable changes in the levels of any of the prominent degradation products of alpha-fetoprotein (a homologue of albumin) mRNA could be observed during this period of development. Since accumulation of degradation intermediates is known to correlate with higher rate of mRNA turnover, our observations raise the possibility that the stability of albumin mRNA may be lower in fetal than in adult mouse liver.
机译:我们已经通过使用横跨白蛋白mRNA中间编码区域的cDNA探针对胎儿小鼠肝脏中的总RNA进行S1核酸酶保护分析,检测到了小鼠白蛋白mRNA的特异性内切核酸酶裂解。使用在其5'末端标记的探针,检测到三个明显的裂解,这些裂解被确认,并通过使用3'末端标记的探针进行进一步分析,确定了它们的内切核酸性质。后者的探针还揭示了另一条切割,这是用5'末端标记的探针未检测到的。这些切割定位于mRNA上的位置,其包括独特的序列基序CCAN1-3CUGN0-1UGAU。始终观察到与这些裂解相对应的降解中间体,特别是在胎儿肝脏中,但在正常或再生的成年肝脏中则没有,并且似乎是在体内起源的。从妊娠第18天开始,它们的水平逐渐降低,到出生后20天才被发现。在此发育期间,未观察到任何甲胎蛋白(白蛋白的同系物)mRNA的显着降解产物水平的可检测变化。由于已知降解中间体的积累与较高的mRNA周转率相关,因此我们的观察结果增加了胎儿白蛋白mRNA稳定性可能低于成年小鼠肝脏的可能性。

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