TY - JOUR
T1 - Plasmodium DEH is ER-localized and crucial for oocyst mitotic division during malaria transmission
AU - Guttery, David S.
AU - Pandey, Rajan
AU - Ferguson, David J. P.
AU - Wall, Richard J.
AU - Brady, Declan
AU - Gupta, Dinesh
AU - Holder, Anthony A.
AU - Tewari, Rita
N1 - Medical Research Council (MRC) Investigator Award and MRC project grants (G0900109, G0900278, and MR/K011782/1)
Newton Bhabha PhD Placement Program (BT/IN/UK/DBT-BC/2015-16)
Cancer Research UK, UK Medical Research Council, Wellcome Trust (FC10097)
PY - 2020/12
Y1 - 2020/12
N2 - Cells use fatty acids (FAs) for membrane biosynthesis, energy storage, and the generation of signaling molecules. 3-hydroxyacyl-CoA dehydratase-DEH-is a key component of very long chain fatty acid synthesis. Here, we further characterized in-depth the location and function of DEH, applying in silico analysis, live cell imaging, reverse genetics, and ultrastructure analysis using the mouse malaria model Plasmodium berghei DEH is evolutionarily conserved across eukaryotes, with a single DEH in Plasmodium spp. and up to three orthologs in the other eukaryotes studied. DEH-GFP live-cell imaging showed strong GFP fluorescence throughout the life-cycle, with areas of localized expression in the cytoplasm and a circular ring pattern around the nucleus that colocalized with ER markers. Δdeh mutants showed a small but significant reduction in oocyst size compared with WT controls from day 10 postinfection onwards, and endomitotic cell division and sporogony were completely ablated, blocking parasite transmission from mosquito to vertebrate host. Ultrastructure analysis confirmed degeneration of Δdeh oocysts, and a complete lack of sporozoite budding. Overall, DEH is evolutionarily conserved, localizes to the ER, and plays a crucial role in sporogony.
AB - Cells use fatty acids (FAs) for membrane biosynthesis, energy storage, and the generation of signaling molecules. 3-hydroxyacyl-CoA dehydratase-DEH-is a key component of very long chain fatty acid synthesis. Here, we further characterized in-depth the location and function of DEH, applying in silico analysis, live cell imaging, reverse genetics, and ultrastructure analysis using the mouse malaria model Plasmodium berghei DEH is evolutionarily conserved across eukaryotes, with a single DEH in Plasmodium spp. and up to three orthologs in the other eukaryotes studied. DEH-GFP live-cell imaging showed strong GFP fluorescence throughout the life-cycle, with areas of localized expression in the cytoplasm and a circular ring pattern around the nucleus that colocalized with ER markers. Δdeh mutants showed a small but significant reduction in oocyst size compared with WT controls from day 10 postinfection onwards, and endomitotic cell division and sporogony were completely ablated, blocking parasite transmission from mosquito to vertebrate host. Ultrastructure analysis confirmed degeneration of Δdeh oocysts, and a complete lack of sporozoite budding. Overall, DEH is evolutionarily conserved, localizes to the ER, and plays a crucial role in sporogony.
UR - http://www.scopus.com/inward/record.url?scp=85094847446&partnerID=8YFLogxK
U2 - 10.26508/lsa.202000879
DO - 10.26508/lsa.202000879
M3 - Article
C2 - 33106323
SN - 2575-1077
VL - 3
JO - Life Science Alliance
JF - Life Science Alliance
IS - 12
M1 - e202000879
ER -
