Classification, expression pattern, and E3 ligase activity assay of rice U-box-containing proteins

Mol Plant. 2008 Sep;1(5):800-15. doi: 10.1093/mp/ssn044.

Abstract

Ubiquitin ligases play a central role in determining the specificity of the ubiquitination system by selecting a myriad of appropriate candidate proteins for modification. The U-box is a recently identified, ubiquitin ligase activity-related protein domain that shows greater presence in plants than in other organisms. In this study, we identified 77 putative U-box proteins from the rice genome using a battery of whole genome analysis algorithms. Most of the U-box protein genes are expressed, as supported by the identification of their corresponding expressed sequence tags (ESTs), full-length cDNAs, or massively parallel signature sequencing (MPSS) tags. Using the same algorithms, we identified 61 U-box proteins from the Arabidopsis genome. The rice and Arabidopsis U-box proteins were classified into nine major classes based on their domain compositions. Comparison between rice and Arabidopsis U-box proteins indicates that the majority of rice and Arabidopsis U-box proteins have the same domain organizations. The inferred phylogeny established the homology between rice and Arabidopsis U-box/ARM proteins. Cell death assay using the rice protoplast system suggests that one rice U-box gene, OsPUB51, might act as a negative regulator of cell death signaling. In addition, the selected U-box proteins were found to be functional E3 ubiquitin ligases. The identification and analysis of rice U-box proteins hereby at the genomic level will help functionally characterize this class of E3 ubiquitin ligase in the future.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Arabidopsis / enzymology
  • Arabidopsis / genetics
  • Biological Assay*
  • Cell Death / genetics
  • Computational Biology
  • Conserved Sequence
  • Gene Expression Regulation, Plant
  • Genes, Plant
  • Genome, Plant
  • Immunity, Innate / genetics
  • Molecular Sequence Data
  • Oryza / enzymology*
  • Oryza / genetics*
  • Phylogeny
  • Plant Proteins / chemistry
  • Plant Proteins / classification*
  • Plant Proteins / genetics*
  • Plant Proteins / metabolism
  • Protein Structure, Tertiary
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction / genetics
  • Ubiquitin-Protein Ligases / chemistry
  • Ubiquitin-Protein Ligases / classification*
  • Ubiquitin-Protein Ligases / genetics*
  • Ubiquitin-Protein Ligases / metabolism

Substances

  • Plant Proteins
  • Ubiquitin-Protein Ligases