Molecular analysis of "de novo" purine biosynthesis in solanaceous species and in Arabidopsis thaliana

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

SN - 1093-9946

VL - 9

SP - 1803

EP - 1816

JO - Frontiers in Bioscience

JF - Frontiers in Bioscience

ER -