Abstract

The ubiquitination pathway regulates growth, development, and stress responses in plants, and the U-box protein family of ubiquitin ligases has important roles in this pathway. Here, 64 putative U-box proteins were identified in the Medicago truncatula genome. In addition to the conserved U-box motif, other functional domains, such as the ARM, kinase, KAP, and WD40 domains, were also detected. Phylogenetic analysis of the M. truncatula U-box proteins grouped them into six subfamilies, and chromosomal mapping and synteny analyses indicated that tandem and segmental duplications may have contributed to the expansion and evolution of the U-box gene family in this species. Using RNA-seq data from M. truncatula seedlings subjected to three different abiotic stresses, we identified 33 stress-inducible plant U-box genes (MtPUBs). Specifically, 25 salinity-, 15 drought-, and 16 cold-regulated MtPUBs were detected. Among them, MtPUB10, MtPUB17, MtPUB18, MtPUB35, MtPUB42, and MtPUB44 responded to all three stress conditions. Expression profiling by qRT-PCR was consistent with the RNA-seq data, and stress-related elements were identified in the promoter regions. The present findings strongly indicate that U-box proteins play critical roles in abiotic stress response in M. truncatula.

Highlights

  • Ubiquitin-mediated proteolysis is required for most cellular processes, and the pathway is mediated by three sequential ubiquitination enzymes, E1, E2, and E3

  • The identified proteins were used for a domain search of the Pfam and SMART databases with an E-value cut-off level of 1.0 or 10, which was recommended for more reliable search results

  • We found 64 proteins containing at least one U-box motif in M. truncatula as annotated by the SMART/Pfam databases, and these proteins were designated as U-box proteins (MtPUB) (Table 1 and S1 Table)

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Summary

Introduction

Ubiquitin-mediated proteolysis is required for most cellular processes, and the pathway is mediated by three sequential ubiquitination enzymes, E1, E2, and E3. E3 ubiquitin ligases are of particular importance as they confer substrate specificity that catalyzes the attachment of ubiquitin to protein targets [1,2]. The E3 ligases can be categorized into distinct families based on their protein domains (RING, HECT, or U-box domains) or mode of action [3,4]. The Ubox E3 ligases, of which there are 64 members in Arabidopsis, were identified most recently and comprise the smallest E3 ligase family [5]. They have an approximately 70-amino-acid conserved U-box motif, which is present in U-box E3 ligases from yeast to humans [6].

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