The Plasmodium falciparum , Nima-related kinase Pfnek-4: a marker for asexual parasites committed to sexual differentiation
Abstract Background Malaria parasites undergo, in the vertebrate host, a developmental switch from asexual replication to sexual differentiation leading to the formation of gametocytes, the only form able to survive in the mosquito vector. Regulation of the onset of the sexual phase remains largely...
Published in: | Malaria Journal |
---|---|
Main Authors: | , , , , |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
BMC
2012
|
Subjects: | |
Online Access: | https://doi.org/10.1186/1475-2875-11-250 https://doaj.org/article/f05c1fc1db294bed93cd791a678ea589 |
id |
ftdoajarticles:oai:doaj.org/article:f05c1fc1db294bed93cd791a678ea589 |
---|---|
record_format |
openpolar |
spelling |
ftdoajarticles:oai:doaj.org/article:f05c1fc1db294bed93cd791a678ea589 2023-05-15T15:15:13+02:00 The Plasmodium falciparum , Nima-related kinase Pfnek-4: a marker for asexual parasites committed to sexual differentiation Reininger Luc Garcia Miguel Tomlins Andrew Müller Sylke Doerig Christian 2012-07-01T00:00:00Z https://doi.org/10.1186/1475-2875-11-250 https://doaj.org/article/f05c1fc1db294bed93cd791a678ea589 EN eng BMC http://www.malariajournal.com/content/11/1/250 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-11-250 1475-2875 https://doaj.org/article/f05c1fc1db294bed93cd791a678ea589 Malaria Journal, Vol 11, Iss 1, p 250 (2012) Gametocytes NIMA Plasmodium falciparum Ser/Thr protein kinase Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2012 ftdoajarticles https://doi.org/10.1186/1475-2875-11-250 2022-12-31T01:41:11Z Abstract Background Malaria parasites undergo, in the vertebrate host, a developmental switch from asexual replication to sexual differentiation leading to the formation of gametocytes, the only form able to survive in the mosquito vector. Regulation of the onset of the sexual phase remains largely unknown and represents an important gap in the understanding of the parasite’s complex biology. Methods The expression and function of the Nima-related kinase Pfnek-4 during the early sexual development of the human malaria parasite Plasmodium falciparum were investigated, using three types of transgenic Plasmodium falciparum 3D7 lines: (i) episomally expressing a Pfnek-4-GFP fusion protein under the control of its cognate pfnek-4 promoter; (ii) episomally expressing negative or positive selectable markers, yeast cytosine deaminase - uridyl phosphoribosyl transferase , or human dihydrofolate reductase , under the control of the pfnek-4 promoter; and (iii) lacking a functional pfnek-4 gene. Parasite transfectants were analysed by fluorescence microscopy and flow cytometry. In vitro growth rate and gametocyte formation were determined by Giemsa-stained blood smears. Results The Pfnek-4-GFP protein was found to be expressed in stage II to V gametocytes and, unexpectedly, in a subset of asexual-stage parasites undergoing schizogony. Culture conditions stimulating gametocyte formation resulted in significant increase of this schizont subpopulation. Moreover, sorted asexual parasites expressing the Pfnek-4-GFP protein displayed elevated gametocyte formation when returned to in vitro culture in presence of fresh red blood cells, when compared to GFP - parasites from the same initial population. Negative selection of asexual parasites expressing pfnek-4 showed a marginal reduction in growth rate, whereas positive selection caused a marked reduction in parasitaemia, but was not sufficient to completely abolish proliferation. Pfnek-4 - clones are not affected in their asexual growth and produced normal numbers of stage V ... Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Malaria Journal 11 1 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Gametocytes NIMA Plasmodium falciparum Ser/Thr protein kinase Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
spellingShingle |
Gametocytes NIMA Plasmodium falciparum Ser/Thr protein kinase Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Reininger Luc Garcia Miguel Tomlins Andrew Müller Sylke Doerig Christian The Plasmodium falciparum , Nima-related kinase Pfnek-4: a marker for asexual parasites committed to sexual differentiation |
topic_facet |
Gametocytes NIMA Plasmodium falciparum Ser/Thr protein kinase Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
Abstract Background Malaria parasites undergo, in the vertebrate host, a developmental switch from asexual replication to sexual differentiation leading to the formation of gametocytes, the only form able to survive in the mosquito vector. Regulation of the onset of the sexual phase remains largely unknown and represents an important gap in the understanding of the parasite’s complex biology. Methods The expression and function of the Nima-related kinase Pfnek-4 during the early sexual development of the human malaria parasite Plasmodium falciparum were investigated, using three types of transgenic Plasmodium falciparum 3D7 lines: (i) episomally expressing a Pfnek-4-GFP fusion protein under the control of its cognate pfnek-4 promoter; (ii) episomally expressing negative or positive selectable markers, yeast cytosine deaminase - uridyl phosphoribosyl transferase , or human dihydrofolate reductase , under the control of the pfnek-4 promoter; and (iii) lacking a functional pfnek-4 gene. Parasite transfectants were analysed by fluorescence microscopy and flow cytometry. In vitro growth rate and gametocyte formation were determined by Giemsa-stained blood smears. Results The Pfnek-4-GFP protein was found to be expressed in stage II to V gametocytes and, unexpectedly, in a subset of asexual-stage parasites undergoing schizogony. Culture conditions stimulating gametocyte formation resulted in significant increase of this schizont subpopulation. Moreover, sorted asexual parasites expressing the Pfnek-4-GFP protein displayed elevated gametocyte formation when returned to in vitro culture in presence of fresh red blood cells, when compared to GFP - parasites from the same initial population. Negative selection of asexual parasites expressing pfnek-4 showed a marginal reduction in growth rate, whereas positive selection caused a marked reduction in parasitaemia, but was not sufficient to completely abolish proliferation. Pfnek-4 - clones are not affected in their asexual growth and produced normal numbers of stage V ... |
format |
Article in Journal/Newspaper |
author |
Reininger Luc Garcia Miguel Tomlins Andrew Müller Sylke Doerig Christian |
author_facet |
Reininger Luc Garcia Miguel Tomlins Andrew Müller Sylke Doerig Christian |
author_sort |
Reininger Luc |
title |
The Plasmodium falciparum , Nima-related kinase Pfnek-4: a marker for asexual parasites committed to sexual differentiation |
title_short |
The Plasmodium falciparum , Nima-related kinase Pfnek-4: a marker for asexual parasites committed to sexual differentiation |
title_full |
The Plasmodium falciparum , Nima-related kinase Pfnek-4: a marker for asexual parasites committed to sexual differentiation |
title_fullStr |
The Plasmodium falciparum , Nima-related kinase Pfnek-4: a marker for asexual parasites committed to sexual differentiation |
title_full_unstemmed |
The Plasmodium falciparum , Nima-related kinase Pfnek-4: a marker for asexual parasites committed to sexual differentiation |
title_sort |
plasmodium falciparum , nima-related kinase pfnek-4: a marker for asexual parasites committed to sexual differentiation |
publisher |
BMC |
publishDate |
2012 |
url |
https://doi.org/10.1186/1475-2875-11-250 https://doaj.org/article/f05c1fc1db294bed93cd791a678ea589 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Malaria Journal, Vol 11, Iss 1, p 250 (2012) |
op_relation |
http://www.malariajournal.com/content/11/1/250 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-11-250 1475-2875 https://doaj.org/article/f05c1fc1db294bed93cd791a678ea589 |
op_doi |
https://doi.org/10.1186/1475-2875-11-250 |
container_title |
Malaria Journal |
container_volume |
11 |
container_issue |
1 |
_version_ |
1766345592044257280 |