The Arabidopsis 14-3-3 protein, GF14omega, binds to the Schizosaccharomyces pombe Cdc25 phosphatase and rescues checkpoint defects in the rad24- mutant

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Sorrell, D. A., Marchbank, A. M., Chrimes, D. A., Dickinson, J. R., Rogers, H. J., Francis, D., Grierson, C. S. and Halford, N. G. 2003. The Arabidopsis 14-3-3 protein, GF14omega, binds to the Schizosaccharomyces pombe Cdc25 phosphatase and rescues checkpoint defects in the rad24- mutant. Planta. 218 (1), pp. 50-57.

AuthorsSorrell, D. A., Marchbank, A. M., Chrimes, D. A., Dickinson, J. R., Rogers, H. J., Francis, D., Grierson, C. S. and Halford, N. G.
Abstract

The fission yeast (S. pombe) mitotic inducer gene, Spcdc25, interacts with the plant cell cycle to establish a small cell size phenotype compared with wild-type cells. We have investigated the nature of this interaction by yeast two-hybrid screening using Spcdc25 as bait in a cDNA library prepared from root tips of Arabidopsis thaliana (L.) Heynh. Three 14-3-3 proteins were detected: G-box Factor-like (GF)14kappa, lambda and omega; binding with Spcdc25 was confirmed by an independent immunoprecipitation assay. To test for cell cycle checkpoint function, GF14kappa, lambda and omega were transformed independently, using the strong nmt1+ promoter, into rad24-, a fission yeast mutant deficient in a 14-3-3 checkpoint protein. When exposed to UV irradiation or in the presence of 10 mM hydroxyurea, only cells transformed with GF14omega could fully rescue the defects in the DNA-damage and DNA-replication checkpoints of this mutant. Supporting evidence for a GF14omega cell cycle function was provided by semi-quantitative reverse transcription-polymerase chain reaction indicating that expression of this gene was elevated in regions of the plant that comprise dividing cells whereas GF14kappa and lambda expression was more evenly detected in all tissues examined. The data are consistent with the hypothesis that interaction between Spcdc25 and the plant cell cycle occurs at the level of a 14-3-3 protein with distinct checkpoint properties.

KeywordsArabidopsis (cell cycle); Cell cycle checkpoints; Schizosaccharomyces; Saccharomyces; 14-3-3 proteins
Year of Publication2003
JournalPlanta
Journal citation218 (1), pp. 50-57
Digital Object Identifier (DOI)doi:10.1007/s00425-003-1083-7
PubMed ID12942327
Open accessPublished as non-open access
FunderBiotechnology and Biological Sciences Research Council
Funder project or code414
521
Metabolic signalling and the partitioning of resources in plants
Output statusPublished
Publication dates
Online26 Aug 2003
Publication process dates
Accepted24 Jun 2003
Copyright licensePublisher copyright
ISSN0032-0935
PublisherSpringer

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