Glycoprotein 46 mRNA abundance is post-transcriptionally regulated during development of Leishmania chagasi promastigotes to an infectious form

Abstract

GP46 is an abundant glycoprotein of 46 kDa on the surface of the promastigote form of most Leishmania species. We show that the steady state level of GP46 mRNA increases >30-fold as Leishmania chagasi promastigotes develop in vitro from a less infectious form during logarithmic growth to a highly infectious form in the stationary phase of cultivation. Nuclear run- on experiments demonstrate that this increase in GP46 mRNA abundance is regulated post-transcriptionally. Plasmids containing the 3'-untranslated regions (UTRs) and downstream intergenic regions (IRs) of two different GP46 genes fused immediately downstream of the β-galactosidase coding region were transfected into L. chagasi, and β-galactosidase activity and mRNA levels were examined. The presence of the 3'-UTR + IR of one GP46 gene (gp46A) resulted in a steady increase in β-galactosidase activity and mRNA level as the transfected promastigotes developed from logarithmic to stationary phase. This differential effect parallels that of the 3'-UTRs + IRs of a family of genes for an unrelated Leishmania surface glycoprotein, GP63. Thus, post- transcriptional regulation of the genes for two different surface glycoproteins of Leishmania occurs via a similar mechanism.