Pf ARID Regulates P. falciparum Malaria Parasite Male Gametogenesis and Female Fertility and Is Critical for Parasite Transmission to the Mosquito Vector

Abstract

Successful completion of the Plasmodium life cycle requires formation of mature gametocytes and their uptake by the female Anopheles mosquito vector in an infected blood meal. Inside the mosquito midgut the parasite undergoes gametogenesis and sexual reproduction.

Plasmodium falciparum CRK5 Is Critical for Male Gametogenesis and Infection of the Mosquito

“…After determining the role of Pf CRK5 in male gametocyte exflagellation, we sought to determine the fertility of individual sex. Since it is not possible to separate male and female gametocytes in in vitro culture, we further analyzed the fertility of male and female Pfcrk5 − gametes utilizing genetic crosses as described previously ( 10 ). For this, we used Pfcrk5 − parasites and transgenic parasite lines, which are sex-sterile for one sex forming either fertile female gametes only ( Pfcdpk4 − ) ( 8 ) or fertile male gametes ( Pfmacfet − ) only ( 23 ), as described previously ( 10 ).…”

“…Since it is not possible to separate male and female gametocytes in in vitro culture, we further analyzed the fertility of male and female Pfcrk5 − gametes utilizing genetic crosses as described previously ( 10 ). For this, we used Pfcrk5 − parasites and transgenic parasite lines, which are sex-sterile for one sex forming either fertile female gametes only ( Pfcdpk4 − ) ( 8 ) or fertile male gametes ( Pfmacfet − ) only ( 23 ), as described previously ( 10 ). WT NF54, Pfcrk5 − , Pfcdpk4 − , and Pfmacfet − gametocytes were generated in vitro in culture for 15 days, and cultures were first fed individually to mosquitoes.…”

“…The P. falciparum NF54 and Pfcrk5 − parasites, asexual and sexual cultures were maintained as described elsewhere ( 10 ). The oligonucleotides used for creation and genotyping analysis of Pfcrk5 − parasites are detailed in Table S1 .…”

“…The oligonucleotides used for creation and genotyping analysis of Pfcrk5 − parasites are detailed in Table S1 . Deletion of PfCRK5 (PlasmoDB identifier Gene - PF3D7_0615500) was achieved as described elsewhere ( 10 ). Two individual clones for Pfcrk5 − (clone 5B and 12F) were used for phenotypic analysis.…”

“…Gametogenesis is further linked to mobilization of intracellular Calcium (Ca 2+ ) via protein kinase G (PKG) ( 5 ), and Calcium-dependent protein kinases (CDPKs), CDPK1 ( 6 ), CDPK2 ( 7 ), and CDPK4 ( 8 ). Several other proteins implicated in this process include a mitogen-activated protein kinase, MAP2 ( 9 ), and an ARID-domain containing protein, Pf ARID ( 10 ).…”