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首页> 外文学位 >Reverse Genetic Analysis of Arabidopsis F-Box Proteins: Role of Auxin-up-Regulated F-Box (AUF) 1 and 2 in Auxin Transport and Cytokinin Signaling.
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Reverse Genetic Analysis of Arabidopsis F-Box Proteins: Role of Auxin-up-Regulated F-Box (AUF) 1 and 2 in Auxin Transport and Cytokinin Signaling.

机译：拟南芥F-Box蛋白的反向遗传分析：生长素上调的F-Box（AUF）1和2在生长素转运和细胞分裂素信号传导中的作用。

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摘要

Selective protein degradation in multicellular organisms is directed by the ubiquitin (Ub) 26S proteasome system (UPS). Through an ATP-dependent enzymatic cascade involving Ub activating enzymes (E1s), Ub conjugation enzymes (E2s) and Ub protein ligases (E3s), Ub becomes covalently attached to various short-lived proteins. The ubiquitylated species are recognized and degraded by the 26S proteasome.;One type of E3s, the SKP1/CULLIN1/F-Box (SCF) ligase, is a four-protein complex which utilizes a F-box protein to determine its substrate specificity. In Arabidopsis thaliana, almost 700 loci have been identified that encode F-box proteins, making this family one of the largest in plants. While a number of F-Box proteins have been shown to control important developmental and physiological processes in plants through forward genetics, the functions of most of them are unknown.;To help define function(s) of the poorly-understood members, I generated a large collection of T-DNA insertion mutants in individual Arabidopsis F-box genes that have not been studied previously. In total, I screened 280 Salk T-DNA insertion alleles and obtained useful mutants for 123 different loci.;Among these F-Box T-DNA insertion mutants, I primarily focused on a set affecting a pair of Auxin-up-Regulated F-Box (AUF) genes, AUF1 and AUF2. AUF1 expression is dramatically induced in the root upon auxin treatment, yet auf1 and auf2 mutants have no significant phenotypic changes in response to auxin. Intriguingly, roots of auf1 and auf1/auf2 plants exhibit increased sensitivity to auxin transport inhibitors. Auxin transport assay reveals that auf1 plants have significant defects in acropetal and basipetal transport in the roots, but the gravitropism response in the auf1 seedlings is normal. In addition, study on the auxin transporter PIN-formed (PIN) proteins revealed that AUF1 regulates polar auxin transport by modulating transcription of multiple PIN genes, which in turn, at least for PIN2, affects protein abundance on the plasma membrane. Roots of auf1 plants also have increased sensitivity to exogenous cytokinin. Root growth kinematic analyses showed that AUF1 does not affect cell division but regulates root cell expansion in the distal elongation zone. Collectively, I conclude that AUF1 is involved in an auxin-cytokinin cross-talk circuit that controls Arabidopsis root development.
机译：多细胞生物中的选择性蛋白质降解由泛素（Ub）26S蛋白酶体系统（UPS）指导。通过涉及Ub活化酶（E1s），Ub缀合酶（E2s）和Ub蛋白连接酶（E3s）的ATP依赖酶级联反应，Ub共价连接到各种短寿命蛋白上。泛素化的物种被26S蛋白酶体识别并降解。一种E3，即SKP1 / CULLIN1 / F-Box（SCF）连接酶，是一种四蛋白复合物，利用F-box蛋白确定其底物特异性。在拟南芥中，已鉴定出将近700个基因座编码F-box蛋白，从而使该家族成为植物中最大的家族之一。尽管已经显示出许多F-Box蛋白通过正向遗传学控制植物中重要的发育和生理过程，但其中大多数功能尚不清楚。为了帮助定义这些成员的功能，我创建了单个拟南芥F-box基因中大量的T-DNA插入突变体，以前没有研究过。总共筛选了280个Salk T-DNA插入等位基因，并获得了123个不同基因座的有用突变体。在这些F-Box T-DNA插入突变体中，我主要关注一组影响一对生长素上调的F-的突变体。 Box（AUF）基因AUF1和AUF2。生长素处理后，根中会明显诱导AUF1表达，但auf1和auf2突变体对生长素的反应没有明显的表型变化。有趣的是，auf1和auf1 / auf2植物的根对生长素运输抑制剂的敏感性增强。生长素转运分析表明，auf1植物在根部的顶基和基部转运方面存在明显缺陷，但auf1幼苗的重力作用是正常的。此外，对生长素转运蛋白PIN形成（PIN）蛋白的研究表明，AUF1通过调节多个PIN基因的转录来调节极性植物生长素的转运，这反过来，至少对于PIN2而言，会影响质膜上的蛋白质丰度。 auf1植物的根对外源细胞分裂素的敏感性也增加了。根生长运动分析表明，AUF1不会影响细胞分裂，但会调节远端延伸区中根细胞的膨胀。总的来说，我得出结论，AUF1参与控制拟南芥根发育的生长素-细胞分裂素串扰电路。

著录项

  • 作者

    Zheng, Xiaohua.;

  • 作者单位

    The University of Wisconsin - Madison.;

  • 授予单位 The University of Wisconsin - Madison.;
  • 学科 Biology Botany.;Biology Genetics.
  • 学位 Ph.D.
  • 年度 2010
  • 页码 207 p.
  • 总页数 207
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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