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赤霉素和脱落酸调节大麦糊粉层的蛋白质合成和 mRNA 水平。

Gibberellic acid and abscisic acid modulate protein synthesis and mRNA levels in barley aleurone layers.

机构信息

Division of Plant Industry, Commonwealth Scientific and Industrial Research Organization, P.O. Box 1600, 2601, Canberra City, A.C.T., Australia.

出版信息

Plant Mol Biol. 1982 Sep;1(3):191-215. doi: 10.1007/BF00021032.

Abstract

Using in vivo pulse labeling, changes in the pattern of protein synthesis were detected in isolated barley aleurone layers treated with fibberellic acid (GA3). GA3 greatly altered the relative rates of synthesis of many polypeptides, increasing some, notably α-amylase, and decreasing others. α-Amylase synthesis increased until it was the major product (over 60%) of protein synthesis after 24h. The pulse-labeled pattern of secreted polypeptides was also changed by GA3. There was the expected increase in α-amylase together with a number of other polypeptides but there was reduced secretion of several polypeptides also.Cell-free translation of RNA isolated from control and hormone-treated tissues was used to measure changes in mRNA levels. GA3 caused many changes, particularly in the level of mRNA for α-amylase. In vitro synthesized α-amylase, identified by immunoaffinity chromatography, had an Mr of 46 000. This polypeptide was partially processed to a polypeptide with Mr 44 000 by the addition of dog pancreas membranes to the in vivo translation mixture. The level of mRNA for α-amylase began to increase 2-4 h after GA3 was added and reached a maximum level of about 20% of total mRNA after 16 h. Thus after 16 h, the synthesis of α-amylase as a proportion of total protein synthesis, continued to increase while the level of its mRNA as a proportion of total mRNA remained constant. These results indicate that protein synthesis was modified more extensively than we can account for by changes in mRNA.Abscisic acid (ABA) reversed all of the effects of GA3 on protein synthesis and mRNA levels. It also promoted synthesis of a small number of new polypeptides and increased the level of some mRNAs. GA3 reversed the accumulation of ABA-promoted mRNAs. Although, ABA strongly suppressed the increase in the level of translatable mRNA for α-amylase, there was an even stronger inhibition of enzyme synthesis and accumulation.We conclude that both GA3 and ABA regulate protein synthesis both positively and negatively in aleurone cells largely by regulating levels of mRNA and in the case of α-amylase, possibly also by changing the efficiency of translation of its mRNA.

摘要

利用活体脉冲标记法,检测到用赤霉素(GA3)处理的大麦糊粉层中蛋白质合成模式的变化。GA3 极大地改变了许多多肽的相对合成速率,增加了一些,特别是α-淀粉酶,而降低了其他多肽的合成速率。α-淀粉酶的合成增加,直到 24 小时后成为蛋白质合成的主要产物(超过 60%)。GA3 还改变了分泌多肽的脉冲标记模式。除了预期的α-淀粉酶增加外,还有许多其他多肽的增加,但也有一些多肽的分泌减少。从对照和激素处理组织中分离的 RNA 进行无细胞翻译,以测量 mRNA 水平的变化。GA3 引起了许多变化,特别是α-淀粉酶的 mRNA 水平。通过免疫亲和层析鉴定的体外合成的α-淀粉酶的 Mr 为 46000。该多肽通过将狗胰腺膜添加到体内翻译混合物中,部分加工成 Mr 为 44000 的多肽。添加 GA3 后 2-4 小时,α-淀粉酶的 mRNA 水平开始增加,16 小时后达到约 20%总 mRNA 的最大值。因此,16 小时后,α-淀粉酶的合成作为总蛋白质合成的比例继续增加,而其作为总 mRNA 的比例保持不变。这些结果表明,蛋白质合成的修饰程度超出了我们可以通过 mRNA 变化来解释的程度。脱落酸(ABA)逆转了 GA3 对蛋白质合成和 mRNA 水平的所有影响。它还促进了少数新多肽的合成,并增加了一些 mRNA 的水平。GA3 逆转了 ABA 促进的 mRNA 的积累。尽管 ABA 强烈抑制了α-淀粉酶可翻译 mRNA 水平的增加,但对酶合成和积累的抑制作用甚至更强。我们得出结论,GA3 和 ABA 主要通过调节 mRNA 水平,在糊粉细胞中正向和负向调节蛋白质合成,并且在α-淀粉酶的情况下,可能还通过改变其 mRNA 的翻译效率来调节。

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