Abstract

Nucleotide synthesis is of central importance to all cells. In most organisms, the purine nucleotides are synthesized de novo from non-nucleotide precursors such as amino acids, ammonia and carbon dioxide. An understanding of the enzymes involved in sugarcane purine synthesis opens the possibility of using these enzymes as targets for chemicals which may be effective in combating phytopathogen. Such an approach has already been applied to several parasites and types of cancer. The strategy described in this paper was applied to identify sugarcane clusters for each step of the de novo purine synthesis pathway. Representative sequences of this pathway were chosen from the National Center for Biotechnology Information (NCBI) database and used to search the translated sugarcane expressed sequence tag (SUCEST) database using the available basic local alignment search tool (BLAST) facility. Retrieved clusters were further tested for the statistical significance of the alignment by an implementation (PRSS3) of the Monte Carlo shuffling algorithm calibrated using known protein sequences of divergent taxa along the phylogenetic tree. The sequences were compared to each other and to the sugarcane clusters selected using BLAST analysis, with the resulting table of p-values indicating the degree of divergence of each enzyme within different taxa and in relation to the sugarcane clusters. The results obtained by this strategy allowed us to identify the sugarcane proteins participating in the purine synthesis pathway.

Highlights

  • The purine nucleotide synthesis and recycling pathways are of central importance to all living organisms (Marr, 1991) because they provide the purines necessary for most biochemical processes such as DNA and RNA metabolism, the biosynthesis intermediates ATP, GTP and coenzymes as well as being an important metabolic regulator in cell signaling (Stryer, 1995)

  • In this paper we describe the identification of the eleven enzymes from the sugarcane expressed sequence (SUCEST) tag project homologous to known de novo purine synthesis proteins

  • The initial analysis of several clusters identified as representatives for each member of the purine synthesis pathway indicated the degree of divergence within those enzymes

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Summary

Introduction

The purine nucleotide synthesis and recycling (or salvage) pathways are of central importance to all living organisms (Marr, 1991) because they provide the purines (adenine and guanine) necessary for most biochemical processes such as DNA and RNA metabolism, the biosynthesis intermediates ATP, GTP and coenzymes as well as being an important metabolic regulator in cell signaling (Stryer, 1995) Due to their importance, purine synthesis pathways have been investigated as potential targets for chemotherapy in several different scenarios (Marr, 1991; Ullman and Carter, 1997; Marr and Ullman, 1995). PRS is an important enzyme involved in the salvage as well as de novo pathway because PRPP is a substrate for the major salvage enzymes adenine-phosphoribosyl-transferase (APRT) and hypoxanthine-guanine-phosphoribosyl-transferase (HGPRT)

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