RRC ID 28121
Author Hooks KB, Turner JE, Graham IA, Runions J, Hooks MA.
Title GFP-tagging of Arabidopsis acyl-activating enzymes raises the issue of peroxisome-chloroplast import competition versus dual localization.
Journal J Plant Physiol
Abstract Protein sequence analysis of a subfamily of 18 Arabidopsis acyl-activating enzymes (AAE) for organelle targeting signals revealed that eight of them possessed putative peroxisomal targeting signals (PTS1), five of which belonged to Clade VI of the AAE superfamily. Peroxisomal localization was confirmed by confocal microscopy of green fluorescent protein (GFP)-AAE fusion proteins co-localizing with peroxisomal RFP. The sequence analysis also revealed that all enzymes of Clade VI possess N-terminal regions indicative of chloroplast transit peptides (cTP). Among the five Clade VI peroxisomal enzymes tested, masking the PTS1 signal with GFP redirected three to plastids. In addition, three other peroxisomal AAEs appeared to be redirected to plastids in AAE-GFP fusion constructs. Due to the lack of evidence supporting plastid localization, we propose that competition dictates the exclusive localization to peroxisomes. AAE2 of Clade VI was the only enzyme with a putative mitochondrial targeting sequence, and it appeared to be targeted to mitochondria. The remainder of the AAEs appeared to be localized to plastids or cytosol. The AAE9-GFP fusion protein appeared to be located within discreet structures within plastids that may be plastoglobules. AAE15-GFP, but not AAE16-GFP appeared to be located in the chloroplast envelope. The number of examples is increasing whereby proteins located within other compartments contribute to plastid function. We provide an example of this through the light-sensitive phenotype of mutants of AAE2.
Volume 169(16)
Pages 1631-8
Published 2012-11-1
DOI 10.1016/j.jplph.2012.05.026
PII S0176-1617(12)00265-9
PMID 22920973
MeSH Amino Acid Sequence Arabidopsis / enzymology* Arabidopsis / genetics Arabidopsis / radiation effects Arabidopsis / ultrastructure Arabidopsis Proteins / metabolism Chloroplasts / enzymology Chloroplasts / metabolism* Coenzyme A Ligases / metabolism* Cytosol / enzymology Cytosol / metabolism Green Fluorescent Proteins Light Luminescent Agents Microscopy, Confocal Mitochondria / enzymology Mitochondria / metabolism* Molecular Sequence Data Mutation Peroxisomes / enzymology Peroxisomes / metabolism* Phenotype Phylogeny Plants, Genetically Modified Protein Transport Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / metabolism Seedlings / enzymology Seedlings / genetics Seedlings / radiation effects Seedlings / ultrastructure Sequence Alignment
IF 3.013
Times Cited 12
WOS Category PLANT SCIENCES
Resource
Arabidopsis / Cultured plant cells, genes pda01713 pda04716 pda06578 pda12997 pda04364 pda10158 pda11718 pda08298 pda06811 pda04511 pda09591 pda12714 pda02888 pdy01839 pdy07652