Plant Molecular Biology ( IF 3.9 ) Pub Date : 2022-05-11 , DOI: 10.1007/s11103-022-01277-6 Kentaro Ezura 1, 2 , Akiyoshi Nakamura 2 , Nobutaka Mitsuda 2
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Key message
Comprehensive yeast and protoplast two-hybrid analyses illustrated the protein–protein interaction network of the TALE homeodomain protein family, KNOX and BLH proteins, in tomato leaf and fruit development.
Abstract
KNOTTED-like (KNOX, KN) proteins and BELL1-like (BLH) proteins, which belong to the same TALE homeodomain family, act together by forming KNOX-BLH heterodimer modules. These modules play crucial roles in regulating multiple developmental processes in plants, like organ differentiation. However, despite the increasing knowledge about individual KNOX and BLH functions, a comprehensive view of their functional protein–protein interaction (PPI) network remains elusive in most plants, including tomato (Solanum lycopersicum), an important model plant to study fruit and leaf development. Here, we characterized eight tomato KNOX genes (SlKN1 to SlKN8) and fourteen tomato BLH genes (SlBLH1 to SlBLH14) by expression profiling, co-expression analysis, and PPI network analysis using two-hybrid techniques in yeasts (Y2H) and protoplasts (P2H). We identified 75 pairwise KNOX-BLH interactions, including ten novel interactors of SlKN2/TKN2, a primary class I KNOX protein, and nine novel interactors of SlKN5, a primary class II KNOX protein. Based on these data, we classified KNOX-BLH modules into several categories, which made us infer the order and combination of the KNOX-BLH modules involved in differentiation processes in leaf and fruit. Notably, the co-expression and interaction of SlKN5 and fruit preferentially expressing BLH1-clade paralogs (SlBLH5/SlBEL11 and SlBLH7) suggest their important roles in regulating fruit differentiation. Furthermore, in silico modeling of the KNOX-BLH modules, sequence analysis, and P2H assay identified several residues and a linker region potentially influencing the affinity of BLHs to KNOXs within their conserved dimerization domains. Together, these findings provide insights into the regulatory mechanism of KNOX-BLH modules underlying tomato organ differentiation.
中文翻译：
番茄中TALE同源域家族和KNOX-BLH相互作用网络的全基因组表征
关键信息
综合酵母和原生质体双杂交分析说明了 TALE 同源域蛋白家族、KNOX 和 BLH 蛋白在番茄叶片和果实发育中的蛋白质相互作用网络。
抽象的
KNOTTED 样 (KNOX, KN) 蛋白和 BELL1 样 (BLH) 蛋白属于同一个 TALE 同源域家族，通过形成 KNOX-BLH 异二聚体模块共同作用。这些模块在调节植物的多个发育过程中起着至关重要的作用，例如器官分化。然而，尽管对单个 KNOX 和 BLH 功能的了解越来越多，但在大多数植物中仍难以全面了解其功能性蛋白质-蛋白质相互作用 (PPI) 网络，包括番茄 ( Solanum lycopersicum )，它是研究果实和叶片发育的重要模式植物. 在这里，我们鉴定了 8 个番茄KNOX基因（SlKN1到SlKN8）和 14 个番茄BLH基因（SlBLH1到SlBLH14) 通过在酵母 (Y2H) 和原生质体 (P2H) 中使用双杂交技术进行表达谱分析、共表达分析和 PPI 网络分析。我们确定了 75 个成对的 KNOX-BLH 相互作用，包括 10 个新的 SlKN2/TKN2 相互作用子（一种初级 I 类 KNOX 蛋白）和 9 个新的 SlKN5 相互作用子（一种初级 II 类 KNOX 蛋白）。基于这些数据，我们将 KNOX-BLH 模块分为几类，这使我们能够推断出参与叶片和果实分化过程的 KNOX-BLH 模块的顺序和组合。值得注意的是，SlKN5 和优先表达 BLH1 进化枝旁系同源物（SlBLH5/SlBEL11 和 SlBLH7）的果实的共表达和相互作用表明它们在调节果实分化中起重要作用。此外，KNOX-BLH 模块的计算机建模、序列分析、和 P2H 分析确定了几个残基和一个接头区域，可能影响 BLH 在其保守的二聚化结构域内对 KNOX 的亲和力。总之，这些发现为番茄器官分化的 KNOX-BLH 模块的调控机制提供了见解。
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