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Approaches to defining dual‐targeted proteins in Arabidopsis
Author(s) -
Carrie Chris,
Kühn Kristina,
Murcha Monika W.,
Duncan Owen,
Small Ian D.,
O’Toole Nicholas,
Whelan James
Publication year - 2009
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/j.1365-313x.2008.03745.x
Subject(s) - arabidopsis , dual (grammatical number) , computational biology , biology , genetics , gene , philosophy , mutant , linguistics
Summary A variety of approaches were used to predict dual‐targeted proteins in Arabidopsis thaliana . These predictions were experimentally tested using GFP fusions. Twelve new dual‐targeted proteins were identified: five that were dual‐targeted to mitochondria and plastids, six that were dual‐targeted to mitochondria and peroxisomes, and one that was dual‐targeted to mitochondria and the nucleus. Two methods to predict dual‐targeted proteins had a high success rate: (1) combining the AraPerox database with a variety of subcellular prediction programs to identify mitochondrial‐ and peroxisomal‐targeted proteins, and (2) using a variety of prediction programs on a biochemical pathway or process known to contain at least one dual‐targeted protein. Several technical parameters need to be taken into account before assigning subcellular localization using GFP fusion proteins. The position of GFP with respect to the tagged polypeptide, the tissue or cells used to detect subcellular localization, and the portion of a candidate protein fused to GFP are all relevant to the expression and targeting of a fusion protein. Testing all gene models for a chromosomal locus is required if more than one model exists.