BACKGROUND: Gametocytes, the blood stages responsible for Plasmodium falciparum transmission, contain electron dense organelles, traditionally named osmiophilic bodies, that are believed to be involved in gamete egress from the host cell. In order to provide novel tools in the cellular and molecular studies of osmiophilic body biology, a P. falciparum transgenic line in which these organelles are specifically marked by a reporter protein was produced and characterized. METHODOLOGY: A P. falciparum transgenic line expressing an 80-residue N-terminal fragment of the osmiophilic body protein Pfg377 fused to the reporter protein DsRed, under the control of pfg377 upstream and downstream regulatory regions, was produced. RESULTS: The transgenic fusion protein is expressed at the appropriate time and stage of sexual differentiation and is trafficked to osmiophilic bodies as the endogenous Pfg377 protein. These results indicate that a relatively small N-terminal portion of Pfg377 is sufficient to target the DsRed reporter to the gametocyte osmiophilic bodies. CONCLUSIONS: This is the first identification of a P. falciparum aminoacid sequence able to mediate trafficking to such organelles. To fluorescently tag such poorly characterized organelles opens novel avenues in cellular and imaging studies on their biogenesis and on their role in gamete egress.
BACKGROUND: Gametocytes, the blood stages responsible for Plasmodium falciparum transmission, contain electron dense organelles, traditionally named osmiophilic bodies, that are believed to be involved in gamete egress from the host cell. In order to provide novel tools in the cellular and molecular studies of osmiophilic body biology, a P. falciparum transgenic line in which these organelles are specifically marked by a reporter protein was produced and characterized. METHODOLOGY: A P. falciparum transgenic line expressing an 80-residue N-terminal fragment of the osmiophilic body protein Pfg377 fused to the reporter protein DsRed, under the control of pfg377 upstream and downstream regulatory regions, was produced. RESULTS: The transgenic fusion protein is expressed at the appropriate time and stage of sexual differentiation and is trafficked to osmiophilic bodies as the endogenous Pfg377 protein. These results indicate that a relatively small N-terminal portion of Pfg377 is sufficient to target the DsRed reporter to the gametocyte osmiophilic bodies. CONCLUSIONS: This is the first identification of a P. falciparum aminoacid sequence able to mediate trafficking to such organelles. To fluorescently tag such poorly characterized organelles opens novel avenues in cellular and imaging studies on their biogenesis and on their role in gamete egress.
Authors: Tomasino Pace; Anna Olivieri; Massimo Sanchez; Veronica Albanesi; Leonardo Picci; Inga Siden Kiamos; Chris J Janse; Andrew P Waters; Elisabetta Pizzi; Marta Ponzi Journal: Mol Microbiol Date: 2006-05 Impact factor: 3.501
Authors: C H Kocken; A M van der Wel; M A Dubbeld; D L Narum; F M van de Rijke; G J van Gemert; X van der Linde; L H Bannister; C Janse; A P Waters; A W Thomas Journal: J Biol Chem Date: 1998-06-12 Impact factor: 5.157
Authors: M Reiss; N Viebig; S Brecht; M N Fourmaux; M Soete; M Di Cristina ; J F Dubremetz; D Soldati Journal: J Cell Biol Date: 2001-02-05 Impact factor: 10.539
Authors: Jessica Kehrer; Mirko Singer; Leandro Lemgruber; Patricia A G C Silva; Friedrich Frischknecht; Gunnar R Mair Journal: PLoS Pathog Date: 2016-07-18 Impact factor: 6.823