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Molecular Biology and Evolution 17:1010-1021 (2000)
© 2000 Society for Molecular Biology and Evolution

Article

Structure and Phylogeny of the Venom Group I Phospholipase A2 Gene

Kandiah Jeyaseelan2,*, Arunmozhiarasi Armugam*, Ma Donghui* and Nget-Hong Tan{dagger}

*Department of Biochemistry, Faculty of Medicine and Bioscience Center, Faculty of Science, National University of Singapore, Singapore; and
{dagger}Department of Biochemistry, Faculty of Medicine, University Malaya, Malaysia

Phospholipases A2 (PLA2s) catalyzing the hydrolysis of phospholipids form a family of proteins with diverse physiological and pharmacological properties. While there have been several reports on the cloning of PLA2 cDNAs, very few studies have been carried out on the PLA2 genes and, most importantly, no information has been available on the gene structure and function of group I venom PLA2. This study, on the PLA2 gene from a spitting cobra, besides being the very first report on any venom group I PLA2 gene, constitutes the missing link in the biology and evolution of phospholipases. The 4-kb gene consists of four exons and three introns and resembles the human pancreatic PLA2 gene. However, the size of intron 3 in particular is much smaller than that in the pancreatic gene. Interestingly, the information for the toxic and most of the pharmacological properties of the venom PLA2 can be attributed to the end of exon 3 and the whole of exon 4 of the gene. This functional delineation fits in well with the theory of adaptive evolution exhibited by the venom PLA2s. We also show that the mammalian pancreatic and elapid PLA2s have similar paths of evolution (probably following gene duplication) from a common ancestral gene. Venom group II phospholipases, although evolved from the same ancestor, diverged early in evolution from the group I PLA2 genes. Intriguingly, CAT reporter gene assays and DNase 1 footprinting studies on the promoter and its deletion constructs using CHO and HepG2 cell lines identified the possible involvement of cis elements such as Sp1, AP2, {gamma}-IRE, and (TG)12 repeats in the expression of the gene in a tissue-specific manner.


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