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Published in: Malaria Journal 1/2012

Open Access 01-12-2012 | Research

Specific tagging of the egress-related osmiophilic bodies in the gametocytes of Plasmodium falciparum

Authors: Anna Rosa Sannella, Anna Olivieri, Lucia Bertuccini, Fabrizio Ferrè, Carlo Severini, Tomasino Pace, Pietro Alano

Published in: Malaria Journal | Issue 1/2012

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Abstract

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.
Appendix
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Literature
1.
go back to reference Kats LM, Cooke BM, Coppel RL, Black CG: Protein trafficking to apical organelles of malaria parasites - building an invasion machine. Traffic. 2008, 9: 176-186.CrossRefPubMed Kats LM, Cooke BM, Coppel RL, Black CG: Protein trafficking to apical organelles of malaria parasites - building an invasion machine. Traffic. 2008, 9: 176-186.CrossRefPubMed
2.
go back to reference Kocken CH, van der Wel AM, Dubbeld MA, Narum DL, van de Rijke FM, van Gemert GJ, van der Linde X, Bannister LH, Janse C, Waters AP, Thomas AW: Precise timing of expression of a Plasmodium falciparum-derived transgene in Plasmodium berghei is a critical determinant of subsequent subcellular localization. J Biol Chem. 1998, 273: 15119-15124. 10.1074/jbc.273.24.15119.CrossRefPubMed Kocken CH, van der Wel AM, Dubbeld MA, Narum DL, van de Rijke FM, van Gemert GJ, van der Linde X, Bannister LH, Janse C, Waters AP, Thomas AW: Precise timing of expression of a Plasmodium falciparum-derived transgene in Plasmodium berghei is a critical determinant of subsequent subcellular localization. J Biol Chem. 1998, 273: 15119-15124. 10.1074/jbc.273.24.15119.CrossRefPubMed
3.
go back to reference Rug M, Wickham ME, Foley M, Cowman AF, Tilley L: Correct promoter control is needed for trafficking of the ring-infected erythrocyte surface antigen to the host cytosol in transfected malaria parasites. Infect Immun. 2004, 72: 6095-6105. 10.1128/IAI.72.10.6095-6105.2004.PubMedCentralCrossRefPubMed Rug M, Wickham ME, Foley M, Cowman AF, Tilley L: Correct promoter control is needed for trafficking of the ring-infected erythrocyte surface antigen to the host cytosol in transfected malaria parasites. Infect Immun. 2004, 72: 6095-6105. 10.1128/IAI.72.10.6095-6105.2004.PubMedCentralCrossRefPubMed
4.
go back to reference Ghoneim A, Kaneko O, Tsuboi T, Torii M: The Plasmodium falciparum RhopH2 promoter and first 24 amino acids are sufficient to target proteins to the rhoptries. Parasitol Int. 2007, 56: 31-43. 10.1016/j.parint.2006.11.001.CrossRefPubMed Ghoneim A, Kaneko O, Tsuboi T, Torii M: The Plasmodium falciparum RhopH2 promoter and first 24 amino acids are sufficient to target proteins to the rhoptries. Parasitol Int. 2007, 56: 31-43. 10.1016/j.parint.2006.11.001.CrossRefPubMed
5.
go back to reference Bannister LH, Margos G, Hopkins JM: Making a home for Plasmodium post-genomics: ultrastructural organisation of the blood stages. Molecular Approaches to Malaria. Edited by: Sherman I. 2005, Washington DC USA: ASM Press Bannister LH, Margos G, Hopkins JM: Making a home for Plasmodium post-genomics: ultrastructural organisation of the blood stages. Molecular Approaches to Malaria. Edited by: Sherman I. 2005, Washington DC USA: ASM Press
6.
go back to reference Hawking F, Wilson ME, Gammage K: Evidence for cyclic development and short-lived maturity in the gametocytes of Plasmodium falciparum. Trans R Soc Trop Med Hyg. 1971, 65: 549-559. 10.1016/0035-9203(71)90036-8.CrossRefPubMed Hawking F, Wilson ME, Gammage K: Evidence for cyclic development and short-lived maturity in the gametocytes of Plasmodium falciparum. Trans R Soc Trop Med Hyg. 1971, 65: 549-559. 10.1016/0035-9203(71)90036-8.CrossRefPubMed
7.
go back to reference Alano P: Molecular approaches to monitor parasite genetic complexity in the transmission of Plasmodium falciparum malaria. Parassitologia. 2005, 47: 199-203.PubMed Alano P: Molecular approaches to monitor parasite genetic complexity in the transmission of Plasmodium falciparum malaria. Parassitologia. 2005, 47: 199-203.PubMed
8.
go back to reference de Koning-Ward TF, Olivieri A, Bertuccini L, Hood A, Silvestrini F, Charvalias K, Berzosa Diaz P, Camarda G, McElwain TF, Papenfuss T, Healer J, Baldassarri L, Crabb BS, Alano P, Ranford-Cartwright LC: The role of osmiophilic bodies and Pfg377 expression in female gametocyte emergence and mosquito infectivity in the human malaria parasite Plasmodium falciparum. Mol Microbiol. 2008, 67: 278-290.CrossRefPubMed de Koning-Ward TF, Olivieri A, Bertuccini L, Hood A, Silvestrini F, Charvalias K, Berzosa Diaz P, Camarda G, McElwain TF, Papenfuss T, Healer J, Baldassarri L, Crabb BS, Alano P, Ranford-Cartwright LC: The role of osmiophilic bodies and Pfg377 expression in female gametocyte emergence and mosquito infectivity in the human malaria parasite Plasmodium falciparum. Mol Microbiol. 2008, 67: 278-290.CrossRefPubMed
9.
go back to reference Ponzi M, Siden-Kiamos I, Bertuccini L, Curra C, Kroeze H, Camarda G, Pace T, Franke-Fayard B, Laurentino EC, Louis C, Waters AP, Janse CJ, Alano P: Egress of Plasmodium berghei gametes from their host erythrocyte is mediated by the MDV-1/PEG3 protein. Cell Microbiol. 2009, 11: 1272-1288. 10.1111/j.1462-5822.2009.01331.x.CrossRefPubMed Ponzi M, Siden-Kiamos I, Bertuccini L, Curra C, Kroeze H, Camarda G, Pace T, Franke-Fayard B, Laurentino EC, Louis C, Waters AP, Janse CJ, Alano P: Egress of Plasmodium berghei gametes from their host erythrocyte is mediated by the MDV-1/PEG3 protein. Cell Microbiol. 2009, 11: 1272-1288. 10.1111/j.1462-5822.2009.01331.x.CrossRefPubMed
10.
go back to reference Talman AM, Lacroix C, Marques SR, Blagborough AM, Carzaniga R, Menard R, Sinden RE: PbGEST mediates malaria transmission to both mosquito and vertebrate host. Mol Microbiol. 2011, 82: 462-474. 10.1111/j.1365-2958.2011.07823.x.CrossRefPubMed Talman AM, Lacroix C, Marques SR, Blagborough AM, Carzaniga R, Menard R, Sinden RE: PbGEST mediates malaria transmission to both mosquito and vertebrate host. Mol Microbiol. 2011, 82: 462-474. 10.1111/j.1365-2958.2011.07823.x.CrossRefPubMed
11.
go back to reference Trager W, Jensen JB: Human malaria parasites in continuous culture. Science. 1976, 193: 673-675. 10.1126/science.781840.CrossRefPubMed Trager W, Jensen JB: Human malaria parasites in continuous culture. Science. 1976, 193: 673-675. 10.1126/science.781840.CrossRefPubMed
12.
go back to reference Fidock DA, Wellems TE: Transformation with human dihydrofolate reductase renders malaria parasites insensitive to WR99210 but does not affect the intrinsic activity of proguanil. Proc Natl Acad Sci USA. 1997, 94: 10931-10936. 10.1073/pnas.94.20.10931.PubMedCentralCrossRefPubMed Fidock DA, Wellems TE: Transformation with human dihydrofolate reductase renders malaria parasites insensitive to WR99210 but does not affect the intrinsic activity of proguanil. Proc Natl Acad Sci USA. 1997, 94: 10931-10936. 10.1073/pnas.94.20.10931.PubMedCentralCrossRefPubMed
13.
go back to reference Gupta SK, Schulman S, Vanderberg JP: Stage-dependent toxicity of N-acetyl-glucosamine to Plasmodium falciparum. J Protozool. 1985, 32: 91-95.CrossRefPubMed Gupta SK, Schulman S, Vanderberg JP: Stage-dependent toxicity of N-acetyl-glucosamine to Plasmodium falciparum. J Protozool. 1985, 32: 91-95.CrossRefPubMed
14.
go back to reference Pace T, Olivieri A, Sanchez M, Albanesi V, Picci L, Siden Kiamos I, Janse CJ, Waters AP, Pizzi E, Ponzi M: Set regulation in asexual and sexual Plasmodium parasites reveals a novel mechanism of stage-specific expression. Mol Microbiol. 2006, 60: 870-882. 10.1111/j.1365-2958.2006.05141.x.CrossRefPubMed Pace T, Olivieri A, Sanchez M, Albanesi V, Picci L, Siden Kiamos I, Janse CJ, Waters AP, Pizzi E, Ponzi M: Set regulation in asexual and sexual Plasmodium parasites reveals a novel mechanism of stage-specific expression. Mol Microbiol. 2006, 60: 870-882. 10.1111/j.1365-2958.2006.05141.x.CrossRefPubMed
15.
go back to reference Lombardo F, Lycett GJ, Lanfrancotti A, Coluzzi M, Arca B: Analysis of apyrase 5' upstream region validates improved Anopheles gambiae transformation technique. BMC Res Notes. 2009, 2: 24-10.1186/1756-0500-2-24.PubMedCentralCrossRefPubMed Lombardo F, Lycett GJ, Lanfrancotti A, Coluzzi M, Arca B: Analysis of apyrase 5' upstream region validates improved Anopheles gambiae transformation technique. BMC Res Notes. 2009, 2: 24-10.1186/1756-0500-2-24.PubMedCentralCrossRefPubMed
16.
go back to reference Newman GR: LR white embedding medium for colloidal gold methods. Colloidal gold - principles, methods and applications. Edited by: Hayat MA. 1989, New York: Academic Press, 2: Newman GR: LR white embedding medium for colloidal gold methods. Colloidal gold - principles, methods and applications. Edited by: Hayat MA. 1989, New York: Academic Press, 2:
17.
go back to reference Saito N, Konishi K, Takeda H, Kato M, Sugiyama T, Asaka M: Antigen retrieval trial for post-embedding immunoelectron microscopy by heating with several unmasking solutions. J Histochem Cytochem. 2003, 51: 989-994. 10.1177/002215540305100802.CrossRefPubMed Saito N, Konishi K, Takeda H, Kato M, Sugiyama T, Asaka M: Antigen retrieval trial for post-embedding immunoelectron microscopy by heating with several unmasking solutions. J Histochem Cytochem. 2003, 51: 989-994. 10.1177/002215540305100802.CrossRefPubMed
18.
go back to reference Alano P, Read D, Bruce M, Aikawa M, Kaido T, Tegoshi T, Bhatti S, Smith DK, Luo C, Hansra S, Carter R, Elliott JF: COS cell expression cloning of Pfg377, a Plasmodium falciparum gametocyte antigen associated with osmiophilic bodies. Mol Biochem Parasitol. 1995, 74: 143-156. 10.1016/0166-6851(95)02491-3.CrossRefPubMed Alano P, Read D, Bruce M, Aikawa M, Kaido T, Tegoshi T, Bhatti S, Smith DK, Luo C, Hansra S, Carter R, Elliott JF: COS cell expression cloning of Pfg377, a Plasmodium falciparum gametocyte antigen associated with osmiophilic bodies. Mol Biochem Parasitol. 1995, 74: 143-156. 10.1016/0166-6851(95)02491-3.CrossRefPubMed
19.
go back to reference Striepen B, Soldati D, Garcia-Reguet N, Dubremetz JF, Roos DS: Targeting of soluble proteins to the rhoptries and micronemes in Toxoplasma gondii. Mol Biochem Parasitol. 2001, 113: 45-53. 10.1016/S0166-6851(00)00379-0.CrossRefPubMed Striepen B, Soldati D, Garcia-Reguet N, Dubremetz JF, Roos DS: Targeting of soluble proteins to the rhoptries and micronemes in Toxoplasma gondii. Mol Biochem Parasitol. 2001, 113: 45-53. 10.1016/S0166-6851(00)00379-0.CrossRefPubMed
20.
go back to reference Olivieri A, Silvestrini F, Sanchez M, Alano P: A 140-bp AT-rich sequence mediates positive and negative transcriptional control of a Plasmodium falciparum developmentally regulated promoter. Int J Parasitol. 2008, 38: 299-312. 10.1016/j.ijpara.2007.08.014.CrossRefPubMed Olivieri A, Silvestrini F, Sanchez M, Alano P: A 140-bp AT-rich sequence mediates positive and negative transcriptional control of a Plasmodium falciparum developmentally regulated promoter. Int J Parasitol. 2008, 38: 299-312. 10.1016/j.ijpara.2007.08.014.CrossRefPubMed
21.
go back to reference Severini C, Silvestrini F, Sannella A, Barca S, Gradoni L, Alano P: The production of the osmiophilic body protein Pfg377 is associated with stage of maturation and sex in Plasmodium falciparum gametocytes. Mol Biochem Parasitol. 1999, 100: 247-252. 10.1016/S0166-6851(99)00050-X.CrossRefPubMed Severini C, Silvestrini F, Sannella A, Barca S, Gradoni L, Alano P: The production of the osmiophilic body protein Pfg377 is associated with stage of maturation and sex in Plasmodium falciparum gametocytes. Mol Biochem Parasitol. 1999, 100: 247-252. 10.1016/S0166-6851(99)00050-X.CrossRefPubMed
22.
go back to reference Lanfrancotti A, Bertuccini L, Silvestrini F, Alano P: Plasmodium falciparum: mRNA co-expression and protein co-localisation of two gene products upregulated in early gametocytes. Exp Parasitol. 2007, 116: 497-503. 10.1016/j.exppara.2007.01.021.CrossRefPubMed Lanfrancotti A, Bertuccini L, Silvestrini F, Alano P: Plasmodium falciparum: mRNA co-expression and protein co-localisation of two gene products upregulated in early gametocytes. Exp Parasitol. 2007, 116: 497-503. 10.1016/j.exppara.2007.01.021.CrossRefPubMed
23.
go back to reference Furuya T, Mu J, Hayton K, Liu A, Duan J, Nkrumah L, Joy DA, Fidock DA, Fujioka H, Vaidya AB, Wellems TE, Su XZ: Disruption of a Plasmodium falciparum gene linked to male sexual development causes early arrest in gametocytogenesis. Proc Natl Acad Sci USA. 2005, 102: 16813-16818. 10.1073/pnas.0501858102.PubMedCentralCrossRefPubMed Furuya T, Mu J, Hayton K, Liu A, Duan J, Nkrumah L, Joy DA, Fidock DA, Fujioka H, Vaidya AB, Wellems TE, Su XZ: Disruption of a Plasmodium falciparum gene linked to male sexual development causes early arrest in gametocytogenesis. Proc Natl Acad Sci USA. 2005, 102: 16813-16818. 10.1073/pnas.0501858102.PubMedCentralCrossRefPubMed
24.
go back to reference Di Cristina M, Spaccapelo R, Soldati D, Bistoni F, Crisanti A: Two conserved amino acid motifs mediate protein targeting to the micronemes of the apicomplexan parasite Toxoplasma gondii. Mol Cell Biol. 2000, 20: 7332-7341. 10.1128/MCB.20.19.7332-7341.2000.PubMedCentralCrossRefPubMed Di Cristina M, Spaccapelo R, Soldati D, Bistoni F, Crisanti A: Two conserved amino acid motifs mediate protein targeting to the micronemes of the apicomplexan parasite Toxoplasma gondii. Mol Cell Biol. 2000, 20: 7332-7341. 10.1128/MCB.20.19.7332-7341.2000.PubMedCentralCrossRefPubMed
25.
go back to reference Reiss M, Viebig N, Brecht S, Fourmaux MN, Soete M, Di Cristina M, Dubremetz JF, Soldati D: Identification and characterization of an escorter for two secretory adhesins in Toxoplasma gondii. J Cell Biol. 2001, 152: 563-578. 10.1083/jcb.152.3.563.PubMedCentralCrossRefPubMed Reiss M, Viebig N, Brecht S, Fourmaux MN, Soete M, Di Cristina M, Dubremetz JF, Soldati D: Identification and characterization of an escorter for two secretory adhesins in Toxoplasma gondii. J Cell Biol. 2001, 152: 563-578. 10.1083/jcb.152.3.563.PubMedCentralCrossRefPubMed
26.
go back to reference Hoppe HC, Ngo HM, Yang M, Joiner KA: Targeting to rhoptry organelles of Toxoplasma gondii involves evolutionarily conserved mechanisms. Nat Cell Biol. 2000, 2: 449-456. 10.1038/35017090.CrossRefPubMed Hoppe HC, Ngo HM, Yang M, Joiner KA: Targeting to rhoptry organelles of Toxoplasma gondii involves evolutionarily conserved mechanisms. Nat Cell Biol. 2000, 2: 449-456. 10.1038/35017090.CrossRefPubMed
27.
go back to reference Bryson K, McGuffin LJ, Marsden RL, Ward JJ, Sodhi JS, Jones DT: Protein structure prediction servers at University College London. Nucleic Acids Res. 2005, W36-W38. 33 Web Server Bryson K, McGuffin LJ, Marsden RL, Ward JJ, Sodhi JS, Jones DT: Protein structure prediction servers at University College London. Nucleic Acids Res. 2005, W36-W38. 33 Web Server
28.
go back to reference Bailey TL, Elkan C: Fitting a mixture model by expectation maximization to discover motifs in biopolymers. Proc Int Conf Intell Syst Mol Biol. 1994, 2: 28-36.PubMed Bailey TL, Elkan C: Fitting a mixture model by expectation maximization to discover motifs in biopolymers. Proc Int Conf Intell Syst Mol Biol. 1994, 2: 28-36.PubMed
29.
go back to reference Grant CE, Bailey TL, Noble WS: FIMO: scanning for occurrences of a given motif. Bioinformatics. 2011, 27 (7): 1017-1018. 10.1093/bioinformatics/btr064.PubMedCentralCrossRefPubMed Grant CE, Bailey TL, Noble WS: FIMO: scanning for occurrences of a given motif. Bioinformatics. 2011, 27 (7): 1017-1018. 10.1093/bioinformatics/btr064.PubMedCentralCrossRefPubMed
31.
go back to reference Camacho C, Coulouris G, Avagyan V, Ma N, Papadopoulos J, Bealer K, Madden TL: BLAST+: architecture and applications. BMC Bioinformatics. 2009, 10: 421-10.1186/1471-2105-10-421.PubMedCentralCrossRefPubMed Camacho C, Coulouris G, Avagyan V, Ma N, Papadopoulos J, Bealer K, Madden TL: BLAST+: architecture and applications. BMC Bioinformatics. 2009, 10: 421-10.1186/1471-2105-10-421.PubMedCentralCrossRefPubMed
32.
go back to reference Petersen TN, Brunak S, von Heijne G, Nielsen H: SignalP 4.0: discriminating signal peptides from transmembrane regions. Nat Methods. 2011, 8 (10): 785-786. 10.1038/nmeth.1701.CrossRefPubMed Petersen TN, Brunak S, von Heijne G, Nielsen H: SignalP 4.0: discriminating signal peptides from transmembrane regions. Nat Methods. 2011, 8 (10): 785-786. 10.1038/nmeth.1701.CrossRefPubMed
33.
go back to reference Gatei W, Kariuki S, Hawley W, ter Kuile F, Terlouw D, Phillips-Howard P, Nahlen B, Gimnig J, Lindblade K, Walker E: Effects of transmission reduction by insecticide-treated bed nets (ITNs) on parasite genetics population structure: I. The genetic diversity of Plasmodium falciparum parasites by microsatellite markers in western Kenya. Malar J. 2010, 9: 353-10.1186/1475-2875-9-353.PubMedCentralCrossRefPubMed Gatei W, Kariuki S, Hawley W, ter Kuile F, Terlouw D, Phillips-Howard P, Nahlen B, Gimnig J, Lindblade K, Walker E: Effects of transmission reduction by insecticide-treated bed nets (ITNs) on parasite genetics population structure: I. The genetic diversity of Plasmodium falciparum parasites by microsatellite markers in western Kenya. Malar J. 2010, 9: 353-10.1186/1475-2875-9-353.PubMedCentralCrossRefPubMed
Metadata
Title
Specific tagging of the egress-related osmiophilic bodies in the gametocytes of Plasmodium falciparum
Authors
Anna Rosa Sannella
Anna Olivieri
Lucia Bertuccini
Fabrizio Ferrè
Carlo Severini
Tomasino Pace
Pietro Alano
Publication date
01-12-2012
Publisher
BioMed Central
Published in
Malaria Journal / Issue 1/2012
Electronic ISSN: 1475-2875
DOI
https://doi.org/10.1186/1475-2875-11-88

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