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JXB Advance Access originally published online on September 12, 2006
Journal of Experimental Botany 2006 57(14):3563-3573; doi:10.1093/jxb/erl107
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© The Author [2006]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

RESEARCH PAPER

A fluoroorotic acid-resistant mutant of Arabidopsis defective in the uptake of uracil

George S. Mourad1,*, Bryan M. Snook1, Joshua T. Prabhakar1, Tyler A. Mansfield1 and Neil P. Schultes2

1Department of Biology, Indiana University-Purdue University Fort Wayne (IPFW), 2101 East Coliseum Blvd, Fort Wayne, IN 46805-1499, USA
2Department of Biochemistry and Genetics, The Connecticut Agricultural Experiment Station (CAES), 123 Huntington Street, New Haven, CT 06511, USA

* To whom correspondence should be addressed. E-mail: mourad{at}ipfw.edu

A fluoroorotic acid (FOA)-resistant mutant of Arabidopsis thaliana was isolated by screening M2 populations of ethyl methane sulphonate (EMS)-mutagenized Columbia seed. FOA resistance was due to a nuclear recessive gene, for1-1, which locates to a 519 kb region in chromosome 5. Assays of key regulatory enzymes in de novo pyrimidine synthesis (uridine monophosphate synthase) and salvage biochemistry (thymidine kinase) confirmed that FOA resistance in for1-1/for1-1 plants was not due to altered enzymatic activities. Uptake studies using radiolabelled purines, pyrimidines, and [14C]FOA reveal that for1-1/for1-1 plants were specifically defective in the uptake of uracil or uracil-like bases. To confirm such specificity, genetic crosses show that FOR1 is a distinct locus from FUR1 which encodes a deoxyuridine nucleoside transporter. In addition, for1-1/for1-1 plants were restored to FOA sensitivity by transformation with the Escherichia coli uracil transporter gene uraA driven by the cauliflower mosaic virus (CaMV) 35S promoter. Molecular mapping studies reveal that FOR1 does not correspond to loci belonging to any of the six known nucleobase transporter families identified in the Arabidopsis genome. Moreover, FOR1 does not appear to regulate the transcript levels of either uracil transporter-encoding loci At2g03590 or At2g03530. The above results strongly suggest that the for1-1 mutant allele affects a transport mechanism that is specific for the uptake of uracil.

Key words: Arabidopsis, fluoroorotic acid resistance, 5-fluorouracil, mutant, pyrimidine, uracil transport


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