Promoter regions of Plasmodium vivax are poorly or not recognized by Plasmodium falciparum

Abstract Background Heterologous promoter analysis in Plasmodium has revealed the existence of conserved cis regulatory elements as promoters from different species can drive expression of reporter genes in heterologous transfection assays. Here, the functional characterization of different Plasmodium vivax promoters in Plasmodium falciparum using luciferase as the reporter gene is presented. Methods Luciferase reporter plasmids harboring the upstream regions of the msp1, dhfr, and vir3 genes as well as the full-length intergenic regions of the vir23/24 and ef-1α genes of P. vivax were constructed and transiently transfected in P. falciparum. Results Only the constructs with the full-length intergenic regions of the vir23/24 and ef-1α genes were recognized by the P. falciparum transcription machinery albeit to values approximately two orders of magnitude lower than those reported by luc plasmids harbouring promoter regions from P. falciparum and Plasmodium berghei. A bioinformatics approach allowed the identification of a motif (GCATAT) in the ef-1α intergenic region that is conserved in five Plasmodium species but is degenerate (GCANAN) in P. vivax. Mutations of this motif in the P. berghei ef-1α promoter region decreased reporter expression indicating it is active in gene expression in Plasmodium. Conclusion Together, this data indicates that promoter regions of P. vivax are poorly or not recognized by the P. falciparum transcription machinery suggesting the existence of P. vivax-specific transcription regulatory elements.

We are grateful to Marcio M. Yamamoto for technical assistance with P. falciparum culture, to Dr. Anamaria A. Camargo for the plasmid bearing the pvmsp1 promoter region, and to Dr. Emilio Fernando Merino and Apuã C.M. Paquola for bioinformatics assistance. To Drs. Alan Cowman, Brendan Crabb and Till Voss for helpful scientific discussions. MFA is a former PhD student from FAPESP (Processo No 01/10653-4). The laboratory of HAP is supported by FAPESP (01/09401-0) and CNPq (ID 302572/2002-3).

We are grateful to Marcio M. Yamamoto for technical assistance with P. falciparum culture, to Dr. Anamaria A. Camargo for the plasmid bearing the pvmsp1 promoter region, and to Dr. Emilio Fernando Merino and Apuã C.M. Paquola for bioinformatics assistance. To Drs. Alan Cowman, Brendan Crabb and Till Voss for helpful scientific discussions. MFA is a former PhD student from FAPESP (Processo No 01/106534). The laboratory of HAP is supported by FAPESP (01/094010) and CNPq (ID 302572/20023).