Signaling networks involving heterotrimeric G-proteins and phospholipids lie at the base of many cellular processes in eukaryotes. Oomycetes possess a family of novel proteins called GPCR-PIPKs (GKs) that are composed of a G-protein-coupled-receptor (GPCR) domain fused to a phosphatidylinositol phosphate kinase (PIPK) domain. Based on this domain structure GKs are anticipated to link G-protein and phospholipid signaling but their functions and biochemical activities are currently unknown. Previously we analyzed the function of one of the twelve GKs in the potato late blight pathogen Phytophthora infestans by gene silencing and overexpression and showed that PiGK4 is involved in spore development, sporangial cleavage, hyphal elongation and virulence (Hua, Meijer et al. 2013, Mol. Microbiology). Overexpression of subdomains of PiGK4 fused to a fluorescent protein revealed that the GPCR domain targets PiGK4 to membranes surrounding certain cellular compartments. With GPCRs as the main target of active agents, and their biological importance, GKs pose potential as oomicide drug targets. To determine the enzymatic activity of the PIPK domain in PiGK4 we make use of the temperature sensitive yeast mutant mss4ts that can be complemented with the fulllength PiGK4 gene. Here we will present a more detailed analysis of the function of the various conserved domains in PiGK4 by complementation essays in yeast mss4ts using modified versions of PiGK4 generated by deleting and swapping domains.
|Title of host publication||Book of Abstracts Oomycete Molecular Genetics Network Meeting|
|Publication status||Published - 2015|
|Event||Oomycete Molecular Genetics Network Meeting 2015 - Pacific Grove, United States|
Duration: 14 Mar 2015 → 17 Mar 2015
|Conference||Oomycete Molecular Genetics Network Meeting 2015|
|Period||14/03/15 → 17/03/15|
van den Hoogen, D. J., Hua, C., Meijer, H. J. G., & Govers, F. (2015). Phytophthora infestans GPCR-PIPK GK4 is a membrane localized PI4P5-kinase and is required for virulence. In Book of Abstracts Oomycete Molecular Genetics Network Meeting (pp. 10)