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腐胺过量产生不会影响杨树和拟南芥中精脒和精胺的分解代谢。

Putrescine overproduction does not affect the catabolism of spermidine and spermine in poplar and Arabidopsis.

机构信息

Department of Biological Sciences, University of New Hampshire, Durham, NH, 03824, USA.

出版信息

Amino Acids. 2014 Mar;46(3):743-57. doi: 10.1007/s00726-013-1581-2. Epub 2013 Sep 8.

Abstract

The effect of up-regulation of putrescine (Put) production by genetic manipulation on the turnover of spermidine (Spd) and spermine (Spm) was investigated in transgenic cells of poplar (Populus nigra × maximowiczii) and seedlings of Arabidopsis thaliana. Several-fold increase in Put production was achieved by expressing a mouse ornithine decarboxylase cDNA either under the control of a constitutive (in poplar) or an inducible (in Arabidopsis) promoter. The transgenic poplar cells produced and accumulated 8-10 times higher amounts of Put than the non-transgenic cells, whereas the Arabidopsis seedlings accumulated up to 40-fold higher amounts of Put; however, in neither case the cellular Spd or Spm increased consistently. The rate of Spd and Spm catabolism and the half-life of cellular Spd and Spm were measured by pulse-chase experiments using [(14)C]Spd or [(14)C]Spm. Spermidine half-life was calculated to be about 22-32 h in poplar and 52-56 h in Arabidopsis. The half-life of cellular Spm was calculated to be approximately 24 h in Arabidopsis and 36-48 h in poplar. Both species were able to convert Spd to Spm and Put, and Spm to Spd and Put. The rates of Spd and Spm catabolism in both species were several-fold slower than those of Put, and the overproduction of Put had only a small effect on the overall rates of turnover of Spd or Spm. There was little effect on the rates of Spd to Spm conversion as well as the conversion of Spm into lower polyamines. While Spm was mainly converted back to Spd and not terminally degraded, Spd was removed from the cells largely through terminal catabolism in both species.

摘要

通过遗传操作上调腐胺(Put)的产生对杨树(Populus nigra × maximowiczii)转基因细胞和拟南芥(Arabidopsis thaliana)幼苗中亚精胺(Spd)和精胺(Spm)周转率的影响进行了研究。通过表达受组成型(在杨树中)或诱导型(在拟南芥中)启动子控制的鼠鸟氨酸脱羧酶 cDNA,使 Put 的产生增加了几倍。转基因杨树细胞产生和积累的 Put 量比非转基因细胞高 8-10 倍,而拟南芥幼苗积累的 Put 量高达 40 倍;然而,在这两种情况下,细胞内 Spd 或 Spm 并没有持续增加。通过使用 [(14)C]Spd 或 [(14)C]Spm 的脉冲追踪实验测量 Spd 和 Spm 的分解代谢率和细胞内 Spd 和 Spm 的半衰期。在杨树中 Spd 的半衰期约为 22-32 h,在拟南芥中为 52-56 h。细胞内 Spm 的半衰期约为 24 h 在拟南芥中,在杨树中为 36-48 h。这两个物种都能够将 Spd 转化为 Spm 和 Put,以及 Spm 转化为 Spd 和 Put。这两个物种中 Spd 和 Spm 的分解代谢率都比 Put 慢几倍,并且 Put 的过度产生对 Spd 或 Spm 的总体周转率的影响很小。对 Spd 向 Spm 的转化以及 Spm 转化为较低的多胺的速率影响很小。虽然 Spm 主要被转化回 Spd 而不是末端降解,但 Spd 主要通过两种物种中的末端分解代谢从细胞中去除。

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