Expression and processing of Plasmodium berghei SERA3 during liver stages.
Cysteine proteases mediate liberation of Plasmodium berghei merozoites from infected hepatocytes. In an attempt to identify the responsible parasite proteases, we screened the genome of P. berghei for cysteine protease-encoding genes. RT-PCR analyses revealed that transcription of four out of five P. berghei serine repeat antigen (PbSERA) genes was strongly upregulated in late liver stages briefly before the parasitophorous vacuole membrane ruptured to release merozoites into the host cell cytoplasm, suggesting a role of PbSERA proteases in these processes. In order to characterize PbSERA3 processing, we raised an antiserum against a non-conserved region of the protein and generated a transgenic P. berghei strain expressing a TAP-tagged PbSERA3 under the control of the endogenous promoter. Immunofluorescence assays revealed that PbSERA3 leaks into the host cell cytoplasm during merozoite development, where it might contribute to host cell death or activate host cell proteases that execute cell death. Importantly, processed PbSERA3 has been detected by Western blot analysis in cell extracts of schizont-infected cells and merozoite-infected detached hepatic cells.
Schmidt-Christensen
Anja
A
Bernhard Nocht Institute for Tropical Medicine, Bernhard-Nocht-Str 74, 20359 Hamburg, Germany.
Sturm
Angelika
A
Horstmann
Sebastian
S
Heussler
Volker T
VT
D000818:Animals; D000953:Antigens, Protozoan / Q000032:analysis / Q000276:immunology* / Q000378:metabolism*; D045744:Cell Line, Tumor; D003546:Cysteine Endopeptidases / Q000032:analysis / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D005260:Female; D008099:Liver / Q000469:parasitology*; D008288:Malaria / Q000276:immunology* / Q000469:parasitology; D030781:Organisms, Genetically Modified; D010962:Plasmodium berghei / Q000254:growth & development / Q000276:immunology* / Q000378:metabolism; D051381:Rats; D015854:Up-Regulation; D014617:Vacuoles / Q000469:parasitology
D000953:Antigens, Protozoan; C057495:serine repeat antigen, Plasmodium; D003546:Cysteine Endopeptidases