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拟南芥的 SKP1 样基因家族在发育过程中表现出高度的差异基因表达和基因产物相互作用。

The SKP1-like gene family of Arabidopsis exhibits a high degree of differential gene expression and gene product interaction during development.

机构信息

Department of Biological Sciences, University of Windsor, Windsor, Ontario, Canada.

出版信息

PLoS One. 2012;7(11):e50984. doi: 10.1371/journal.pone.0050984. Epub 2012 Nov 30.

Abstract

The Arabidopsis thaliana genome encodes several families of polypeptides that are known or predicted to participate in the formation of the SCF-class of E3-ubiquitin ligase complexes. One such gene family encodes the Skp1-like class of polypeptide subunits, where 21 genes have been identified and are known to be expressed in Arabidopsis. Phylogenetic analysis based on deduced polypeptide sequence organizes the family of ASK proteins into 7 clades. The complexity of the ASK gene family, together with the close structural similarity among its members raises the prospect of significant functional redundancy among select paralogs. We have assessed the potential for functional redundancy within the ASK gene family by analyzing an expanded set of criteria that define redundancy with higher resolution. The criteria used include quantitative expression of locus-specific transcripts using qRT-PCR, assessment of the sub-cellular localization of individual ASK:YFP auto-fluorescent fusion proteins expressed in vivo as well as the in planta assessment of individual ASK-F-Box protein interactions using bimolecular fluorescent complementation techniques in combination with confocal imagery in live cells. The results indicate significant functional divergence of steady state transcript abundance and protein-protein interaction specificity involving ASK proteins in a pattern that is poorly predicted by sequence-based phylogeny. The information emerging from this and related studies will prove important for defining the functional intersection of expression, localization and gene product interaction that better predicts the formation of discrete SCF complexes, as a prelude to investigating their molecular mode of action.

摘要

拟南芥基因组编码了几个家族的多肽,这些多肽已知或预测参与形成 SCF 类 E3 泛素连接酶复合物。这样的基因家族之一编码 Skp1 样多肽亚基家族,其中已鉴定出 21 个基因在拟南芥中表达。基于推导的多肽序列的系统发育分析将 ASK 蛋白家族组织成 7 个分支。ASK 基因家族的复杂性,以及其成员之间的紧密结构相似性,提出了选择的旁系同源物之间存在显著功能冗余的可能性。我们通过分析定义更高分辨率冗余的扩展标准集来评估 ASK 基因家族内的潜在功能冗余。使用的标准包括使用 qRT-PCR 定量表达基因座特异性转录物、评估个体 ASK:YFP 自体荧光融合蛋白在体内的亚细胞定位,以及使用双分子荧光互补技术结合共焦成像在活细胞中评估个体 ASK-F-Box 蛋白相互作用。结果表明,ASK 蛋白的稳定状态转录物丰度和蛋白质-蛋白质相互作用特异性存在显著的功能分化,其模式与基于序列的系统发育预测的模式相差很大。这些信息以及相关研究的结果对于定义表达、定位和基因产物相互作用的功能交集将非常重要,这可以更好地预测离散 SCF 复合物的形成,作为研究其分子作用模式的前奏。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6e44/3511428/61a824cc7fa6/pone.0050984.g001.jpg

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