| Successful modification | The parasite was generated by the genetic modification |
| The mutant contains the following genetic modification(s) |
Gene tagging
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| Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 24594931
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| MR4 number |
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| Parent parasite used to introduce the genetic modification |
| Rodent Malaria Parasite | P. berghei |
| Parent strain/line | P. berghei ANKA |
| Name parent line/clone |
P. berghei ANKA 2.34
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| Other information parent line | P. berghei ANKA 2.34 is a cloned, gametocyte producer line of the ANKA strain (PubMed: PMID: 15137943). |
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| The mutant parasite was generated by |
| Name PI/Researcher | Brochet, M; Billker, O. |
| Name Group/Department | Wellcome Trust Sanger Institute, Hinxton |
| Name Institute | Wellcome Trust Sanger Institute, Hinxton |
| City | Hinxton, Cambridge |
| Country | UK |
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| Name of the mutant parasite |
| RMgm number | RMgm-998 |
| Principal name | PIP5K-HA |
| Alternative name | |
| Standardized name | |
| Is the mutant parasite cloned after genetic modification | No |
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| Phenotype |
| Asexual blood stage | Not tested |
| Gametocyte/Gamete | Not tested |
| Fertilization and ookinete | Expression of PIP5K-HA in ookinetes |
| Oocyst | Not tested |
| Sporozoite | Not tested |
| Liver stage | Not tested |
| Additional remarks phenotype | Mutant/mutation
The mutant expressses a C-terminal HA-tagged version of PIP5K
Protein (function)
Enzymes in the inositol phospholipid biosynthetic pathway. Phosphorylated phosphatidylinositol lipids have important roles in vesicle trafficking and as a source of secondary messengers in signal transduction. Their biosynthesis from phosphatidyl-1D-myo-inositol (PI) is mediated by lipid kinases. The P. berghei genome encodes four putative lipid kinases to convert PI first to phosphatidylinositol 4-phosphate (PI4P) and then to phosphatidylinositol (4,5)- bisphosphate (PI(4,5)P2). Hydrolysis of the latter by a PI-specific phospholipase C (PI-PLC) gives rise to the secondary messenger inositol (1,4,5)-trisphosphate (IP3), which plays an important role in P. berghei gametocytes, where it is responsible for the mobilisation of Ca2+ from internal stores, leading to activation and gametogenesis.
To test whether the PKG-dependent phosphorylation of enzymes associated with phosphoinositide metabolism has a direct role in ookinete motility, an experimental genetics approach was used to infer the role of putative PI kinases. Both the putative PI4K (PBANKA_110940) and the putative PIP5K (PBANKA_020310) were unable to be genetically disrupted (unpublished data), suggesting these genes may be essential for asexual growth, although both loci could be modified.
Phenotype
Expression of PIP5K-HA in ookinetes
Additional information
The location of PIP5K-HA rapidly redistributed from the cell periphery to the ookinete cytosol in ookinetes when treated with 0.5 mM C2 (an inhibitor of PKG), supporting a direct link between PKG and phosphoinositide metabolism.
Other mutants
RMgm-997: Unsuccessful attempts to disrupt PIP5K
RMgm-996: Unsuccessful attempts to disrupt PI4K (PBANKA_110940)
RMgm-969: A mutant expressing a mutated form of PI4K (PBANKA_110940)
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*RMgm-998
