Additional remarks phenotype | Mutant/mutation
The mutant (Pla1mch) expresses a C-terminal 3XHA-mCherry tagged version of of Pla1.
Protein (function)
A total of 22 genes have been identified thus far, having similarities to the representative members of at least one of the four groups of known phospholipases; PLA, PLB, PLC and PLD. Out of these, 15 have the α/β hydrolase fold, four have the patatin-like phospholipase (PLP) domain and the remaining three have lecithin:cholesterol acyltransferase (LCAT), exonuclease-endonuclease-phosphatase (EEP) and DDHD domains. An interesting class of enzymes is the intracellular PLA1 family. In most eukaryotes, there is only one enzyme of this family while there are three members in mammals. Early characterization efforts identified a phosphatidic acid (PA) preferring phospholipase A1 (PAPLA1) a putative phospholipase, DDHD1 (PbPla1, PlasmoDB accession number: PBANKA_1423100), which is the only P. berghei homolog of PA-PLA1 (Pla1)
Phenotype
mCherry fluorescence was detected in the parasite’s blood stages indicating the presence of PbPla1 protein during the entire erythrocytic cycle. Protein expression of Pla1 representing the correct size of the fusion protein was confirmed by western blot. No fluorescence could be detected in oocyst or salivary gland sporozoites. Expression of Pla1 protein was maintained during the mixed blood stage (MBS) and early to mid-liver stages. Protein expression is maintained during late EEF development and is also detected in detached cells marking the end of the hepatic phase. Further analysis indicated that PbPla1 is not associated with either the PVM or the merozoite plasma membrane as is evident by its co-staining with anti-UIS4 and anti-MSP1.
Additional information
See mutant RMgm-5203 lacking expression of Pla1, showing evidence for a role in liver stage merosome release
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