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马铃薯ADP-葡萄糖焦磷酸化酶小亚基的结构与表达

Structure and expression of the potato ADP-glucose pyrophosphorylase small subunit.

作者信息

Nakata P A, Anderson J M, Okita T W

机构信息

Department of Biochemistry and Biophysics, Washington State University, Pullman 99164-6340.

出版信息

J Biol Chem. 1994 Dec 9;269(49):30798-807.

PMID:7983010
Abstract

ADP-glucose pyrophosphorylase (AGP) catalyzes a key regulatory step in starch synthesis. To elucidate the molecular basis for the expression of the potato (Solanum tuberosum L.) AGP during tuber development, the structure of the small subunit AGP (sAGP) gene and its patterns of expression were examined. DNA sequence analysis revealed that the sAGP gene is over 5.5 kilobases long and has a complex structure including eight introns. Unlike the situation in other plants where tissue-specific sAGP are found, our Southern and Northern blot analysis indicated that the same sAGP gene is expressed both in tubers (non-photosynthetic tissue) and leaves (photosynthetic tissue). These data were supported by comparing sequences of isolated sAGP leaf cDNAs to the tuber cDNA sequence, by primer extension analysis of leaf and tuber poly(A)+ RNAs, and by the spatial expression patterns of a gusA (beta-glucuronidase) reporter gene driven by the potato sAGP promoter in transgenic potato plants. Although the sAGP gene appeared to be transcriptionally controlled in both developing tubers and in leaves, the relative level of leaf antigen was significantly lower than its level of transcript, indicating that sAGP expression in leaves is primarily regulated post-transcriptionally. The observed tissue type-dependent regulation of sAGP expression appears to control the extent of starch biosynthesis by regulating the levels of this enzyme and, thus, alleviate the need for tissue-specific forms of the sAGP in potato.

摘要

ADP - 葡萄糖焦磷酸化酶(AGP)催化淀粉合成中的关键调控步骤。为阐明马铃薯(Solanum tuberosum L.)AGP在块茎发育过程中表达的分子基础,对小亚基AGP(sAGP)基因的结构及其表达模式进行了研究。DNA序列分析表明,sAGP基因长度超过5.5千碱基,具有复杂的结构,包括8个内含子。与在其他植物中发现组织特异性sAGP的情况不同,我们的Southern和Northern印迹分析表明,同一个sAGP基因在块茎(非光合组织)和叶片(光合组织)中均有表达。通过比较分离的sAGP叶片cDNA序列与块茎cDNA序列、对叶片和块茎poly(A)+ RNA进行引物延伸分析以及由马铃薯sAGP启动子驱动的gusA(β - 葡萄糖醛酸酶)报告基因在转基因马铃薯植株中的空间表达模式,这些数据得到了支持。尽管sAGP基因在发育中的块茎和叶片中似乎都受到转录调控,但叶片抗原的相对水平明显低于其转录水平,这表明叶片中sAGP的表达主要在转录后受到调控。观察到的sAGP表达的组织类型依赖性调控似乎通过调节该酶的水平来控制淀粉生物合成的程度,从而减少了马铃薯中对sAGP组织特异性形式的需求。

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