A specific feature of the angiosperm Rab escort protein (REP) and evolution of the REP/GDI superfamily
Language English Country Netherlands Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S., Research Support, U.S. Gov't, P.H.S.
PubMed
15854662
DOI
10.1016/j.jmb.2005.02.002
PII: S0022-2836(05)00142-7
Knihovny.cz E-resources
- MeSH
- Adaptor Proteins, Signal Transducing MeSH
- Alkyl and Aryl Transferases classification genetics physiology MeSH
- Arabidopsis genetics metabolism MeSH
- Point Mutation genetics MeSH
- Cytoplasm chemistry MeSH
- Phylogeny MeSH
- Guanine Nucleotide Dissociation Inhibitors classification genetics metabolism MeSH
- Cloning, Molecular MeSH
- Conserved Sequence MeSH
- Evolution, Molecular * MeSH
- Molecular Sequence Data MeSH
- Protein Prenylation genetics physiology MeSH
- Arabidopsis Proteins analysis genetics physiology MeSH
- rab GTP-Binding Proteins genetics metabolism MeSH
- Saccharomyces cerevisiae Proteins genetics metabolism MeSH
- Saccharomyces cerevisiae genetics MeSH
- Amino Acid Sequence MeSH
- Genetic Complementation Test MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Research Support, U.S. Gov't, P.H.S. MeSH
- Names of Substances
- Adaptor Proteins, Signal Transducing MeSH
- Alkyl and Aryl Transferases MeSH
- Guanine Nucleotide Dissociation Inhibitors MeSH
- MRS6 protein, S cerevisiae MeSH Browser
- Arabidopsis Proteins MeSH
- Rab escort protein, Arabidopsis MeSH Browser
- rab GTP-Binding Proteins MeSH
- Saccharomyces cerevisiae Proteins MeSH
Rab GTPases participating in the regulation of vesicle trafficking in eukaryotes are geranylgeranylated by the Rab geranylgeranyl transferase (RabGGTase) in complex with the Rab escort protein (REP). Here, we describe basic properties of the Arabidopsis thaliana REP (AthREP), first REP outside yeasts or metazoans to be characterized. GFP-tagged AthREP, as well as the geranylgeranylation activity, were localized predominantly to the cytoplasm. Recombinant AthREP interacted with yeast 6His-Ypt1, tobacco 6His-RabA1a, and Arabidopsis RabA2a in vitro preferring the GDP-bound form of the latter. Recombinant AthREP with C-terminal but not N-terminal tags stimulated geranylgeranylation of various Rab GTPases in Arabidopsis extracts in vitro. Neither recombinant AthREP protein exhibited activity in yeast extracts, while recombinant yeast REP (6His-SceMrs6) stimulated Rab geranylgeranylation in all extracts tested. We found that a conserved arginine residue, R195, known to be crucial for yeast REP function, is substituted by an asparagine or threonine residue in angiosperm REPs. A point mutant allele of AthREP with arginine at this position complemented the yeast REP mutation, while wild-type AthREP did not. Based on phylogenetic analysis of REP and GDP dissociation inhibitor (GDI) sequences from a broad range of eukaryotic lineages, we propose a new view on evolution of the REP/GDI superfamily with a bi-functional REP/GDI protein as a direct ancestor.
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Protein Prenylation in Plant Stress Responses
