RESUMO
All Plasmodium species express variant antigens which may mediate immune escape in the vertebrate host. In Plasmodium falciparum, the rif gene family encodes variant antigens which are partly exposed on the infected red blood cell surface and may function as virulence factors. Not all rif genes are expressed at the same time and it is unclear what controls rif gene expression. In this work, we addressed global rif transcription using plasmid vectors with two drug resistance markers, one controlled by a rif 5' upstream region and the second by a constitutively active promoter. After spontaneous integration into the genome of one construct, we observed that the resistance marker controlled by the rif 5' upstream region was expressed dependent on the applied drug pressure. Then, the global transcription of rif genes in these transfectants was compared in the presence or absence of drugs. The relative transcript quantities of all rif loci did not change profoundly between strains grown with or without drug. We conclude that either there is no crosstalk between rif loci or that the elusive system of allelic exclusion of rif gene transcription is not controlled by their 5' upstream region alone.
Assuntos
Genoma de Protozoário/genética , Plasmodium falciparum/genética , Regiões Promotoras Genéticas/genética , Antígenos de Protozoários/genética , Antígenos de Protozoários/imunologia , Plasmodium falciparum/imunologia , Proteínas de Protozoários/genética , Proteínas de Protozoários/imunologiaRESUMO
The genome of the malaria parasite Plasmodium falciparum contains the surf gene family which encodes large transmembrane proteins of unknown function. While some surf alleles appear to be expressed in sexual stages, others occur in asexual blood stage forms and may be associated to virulence-associated processes and undergo transcriptional switching. We accessed the transcription of surf genes along multiple invasions by real time PCR. Based on the observation of persistent expression of gene surf4.1, we created a parasite line which expresses a conditionally destabilized SURFIN4.1 protein. Upon destabilization of the protein, no interference of parasite growth or morphological changes were detected. However, we observed a strong increase in the transcript quantities of surf4.1 and sometimes of other surf genes in knocked-down parasites. While this effect was reversible when SURFIN4.1 was stabilized again after a few days of destabilization, longer destabilization periods resulted in a transcriptional switch away from surf4.1. When we tested if a longer transcript half-life was responsible for increased transcript detection in SURFIN4.1 knocked-down parasites, no alteration was found compared to control parasite lines. This suggests a specific feedback of the expressed SURFIN protein to its transcript pointing to a novel type of regulation, inedited in Plasmodium.