Molecular cloning and expression pattern of 14‐3‐3ζ from the malaria vector, Anopheles sinensis

Abstract

Abstract14‐3‐3 proteins are known to play a pivotal role in cell survival, apoptosis and signal transduction. The 14‐3‐3ζ isoform has been cloned and characterized from many eukaryotic organisms, including the fruit fly and silkworm. However, no study on mosquito 14‐3‐3 has been reported to date. In an attempt to investigate the function of 14‐3‐3 in midgut epithelial cells undergoing apoptosis, a cDNA library was generated from the malaria vector, Anopheles sinensis, which was treated with apoptosis‐inducing Actinomycin‐D. We were able to identify and obtain A. sinensis 14‐3‐3ζ cDNA (Ansi14‐3‐3ζ) from expressed sequence tags (EST) analysis after conducting massive sequencing of the A. sinensis cDNA library. Ansi14‐3‐3ζ has very high homology to 14‐3‐3 homologs of various insects, such as Anopheles gambiae (100%), Aedes aegypti (100%), Drosophila melanogaster (96%), Bombyx mori (93%), Apis mellifera (93%) and Mus musculus (81%), indicating that mosquito 14‐3‐3ζ is a highly conserved gene in diverse organisms. Analysis of temporal expression patterns showed that Ansi14‐3‐3ζ mRNA is highly expressed in egg, early pupae and adult stages and is also expressed, although at low levels, in fourth instar larvae and late pupae. In response to two immune elicitors (lipopolysaccharide and laminarin), no striking induction of 14‐3‐3ζ mRNA was observed in A. sinensis. Further studies of the precise biological function, inducibility and subcellular distribution of 14‐3‐3ζ are required in Plasmodium invasion‐induced apoptotic midgut cells in A. sinensis in the context of the Time Bomb model.