Molecular analysis of "de novo" purine biosynthesis in solanaceous species and in Arabidopsis thaliana
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Molecular analysis of "de novo" purine biosynthesis in solanaceous species and in Arabidopsis thaliana. / van der Graaff, Eric; Hooykaas, Paul; Lein, Wolfgang; Lerchl, Jens; Kunze, Gotthard; Sonnewald, Uwe; Boldt, Ralf.
I: Frontiers in Bioscience, Bind 9, 01.05.2004, s. 1803-16.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning
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TY - JOUR
T1 - Molecular analysis of "de novo" purine biosynthesis in solanaceous species and in Arabidopsis thaliana
AU - van der Graaff, Eric
AU - Hooykaas, Paul
AU - Lein, Wolfgang
AU - Lerchl, Jens
AU - Kunze, Gotthard
AU - Sonnewald, Uwe
AU - Boldt, Ralf
PY - 2004/5/1
Y1 - 2004/5/1
N2 - Purine nucleotides are essential components to sustain plant growth and development. In plants they are either synthesized "de novo" during the process of purine biosynthesis or are recycled from purine bases and purine nucleosides throughout the salvage pathway. Comparison between animals, microorganisms and Arabidopsis, the first plant species with a completely sequenced genome, shows that plants principally use the same biochemical steps to synthesize purine nucleotides and possess all the essential genes and enzymes. Here we report on the cloning and molecular analysis of the complete purine biosynthesis pathway in plants, and the in planta functional analysis of PRPP (5-phosphoribosyl-1-pyrophoshate) amidotransferase (ATase), catalyzing the first committed step of the "de novo" purine biosynthesis. The cloning of the genes involved in the purine biosynthesis pathway was attained by a screening strategy with heterologous cDNA probes and by using S. cerevisiae mutants for complementation. Southern hybridization showed a complex genomic organization for these genes in solanaceous species and their organ- and developmental specific expression was analyzed by Northern hybridization. The specific role of ATase for plant growth and development was analyzed in transgenic tobacco plants exhibiting a reduced ATase activity and in an Arabidopsis T-DNA mutant (atd2) deficient for ATase2. The transgenic tobacco plants as well as the Arabidopsis mutant exhibit a specific and comparable phenotype, which is characterized by strong growth retardation and severe chlorosis in leaves. The formation of white leaves, but green cotyledons is a characteristic trait of the Arabidopsis atd2 mutant.
AB - Purine nucleotides are essential components to sustain plant growth and development. In plants they are either synthesized "de novo" during the process of purine biosynthesis or are recycled from purine bases and purine nucleosides throughout the salvage pathway. Comparison between animals, microorganisms and Arabidopsis, the first plant species with a completely sequenced genome, shows that plants principally use the same biochemical steps to synthesize purine nucleotides and possess all the essential genes and enzymes. Here we report on the cloning and molecular analysis of the complete purine biosynthesis pathway in plants, and the in planta functional analysis of PRPP (5-phosphoribosyl-1-pyrophoshate) amidotransferase (ATase), catalyzing the first committed step of the "de novo" purine biosynthesis. The cloning of the genes involved in the purine biosynthesis pathway was attained by a screening strategy with heterologous cDNA probes and by using S. cerevisiae mutants for complementation. Southern hybridization showed a complex genomic organization for these genes in solanaceous species and their organ- and developmental specific expression was analyzed by Northern hybridization. The specific role of ATase for plant growth and development was analyzed in transgenic tobacco plants exhibiting a reduced ATase activity and in an Arabidopsis T-DNA mutant (atd2) deficient for ATase2. The transgenic tobacco plants as well as the Arabidopsis mutant exhibit a specific and comparable phenotype, which is characterized by strong growth retardation and severe chlorosis in leaves. The formation of white leaves, but green cotyledons is a characteristic trait of the Arabidopsis atd2 mutant.
KW - Amidophosphoribosyltransferase
KW - Arabidopsis
KW - Blotting, Northern
KW - Cloning, Molecular
KW - Gene Expression
KW - Genes, Plant
KW - Plants, Genetically Modified
KW - Purines
KW - Solanaceae
KW - Solanum tuberosum
KW - Tobacco
M3 - Journal article
C2 - 14977588
VL - 9
SP - 1803
EP - 1816
JO - Frontiers in Bioscience
JF - Frontiers in Bioscience
SN - 1093-9946
ER -
ID: 106391467