Characterization of GEXP15 as a Potential Regulator of Protein Phosphatase 1 in <i>Plasmodium falciparum</i>
The Protein Phosphatase type 1 catalytic subunit (PP1c) (PF3D7_1414400) operates in combination with various regulatory proteins to specifically direct and control its phosphatase activity. However, there is little information about this phosphatase and its regulators in the human malaria parasite,...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-08-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/24/16/12647 |
| _version_ | 1797584524221087744 |
|---|---|
| author | Hala Mansour Alejandro Cabezas-Cruz Véronique Peucelle Amaury Farce Sophie Salomé-Desnoulez Ines Metatla Ida Chiara Guerrera Thomas Hollin Jamal Khalife |
| author_facet | Hala Mansour Alejandro Cabezas-Cruz Véronique Peucelle Amaury Farce Sophie Salomé-Desnoulez Ines Metatla Ida Chiara Guerrera Thomas Hollin Jamal Khalife |
| author_sort | Hala Mansour |
| collection | DOAJ |
| description | The Protein Phosphatase type 1 catalytic subunit (PP1c) (PF3D7_1414400) operates in combination with various regulatory proteins to specifically direct and control its phosphatase activity. However, there is little information about this phosphatase and its regulators in the human malaria parasite, <i>Plasmodium falciparum</i>. To address this knowledge gap, we conducted a comprehensive investigation into the structural and functional characteristics of a conserved <i>Plasmodium</i>-specific regulator called Gametocyte EXported Protein 15, GEXP15 (PF3D7_1031600). Through in silico analysis, we identified three significant regions of interest in GEXP15: an N-terminal region housing a PP1-interacting RVxF motif, a conserved domain whose function is unknown, and a GYF-like domain that potentially facilitates specific protein–protein interactions. To further elucidate the role of GEXP15, we conducted in vitro interaction studies that demonstrated a direct interaction between GEXP15 and PP1 via the RVxF-binding motif. This interaction was found to enhance the phosphatase activity of PP1. Additionally, utilizing a transgenic GEXP15-tagged line and live microscopy, we observed high expression of GEXP15 in late asexual stages of the parasite, with localization predominantly in the nucleus. Immunoprecipitation assays followed by mass spectrometry analyses revealed the interaction of GEXP15 with ribosomal- and RNA-binding proteins. Furthermore, through pull-down analyses of recombinant functional domains of His-tagged GEXP15, we confirmed its binding to the ribosomal complex via the GYF domain. Collectively, our study sheds light on the PfGEXP15–PP1–ribosome interaction, which plays a crucial role in protein translation. These findings suggest that PfGEXP15 could serve as a potential target for the development of malaria drugs. |
| first_indexed | 2024-03-10T23:52:56Z |
| format | Article |
| id | doaj.art-f2cb222fbdee41989b2f496902abc253 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T23:52:56Z |
| publishDate | 2023-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-f2cb222fbdee41989b2f496902abc2532023-11-19T01:26:50ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-08-0124161264710.3390/ijms241612647Characterization of GEXP15 as a Potential Regulator of Protein Phosphatase 1 in <i>Plasmodium falciparum</i>Hala Mansour0Alejandro Cabezas-Cruz1Véronique Peucelle2Amaury Farce3Sophie Salomé-Desnoulez4Ines Metatla5Ida Chiara Guerrera6Thomas Hollin7Jamal Khalife8Univ. Lille, CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, U1019-UMR 9017-CIIL-Center for Infection and Immunity of Lille, 59000 Lille, FranceANSES, INRAE, Ecole Nationale Vétérinaire d’Alfort, UMR BIPAR, Laboratoire de Santé Animale, 94700 Maisons-Alfort, FranceUniv. Lille, CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, U1019-UMR 9017-CIIL-Center for Infection and Immunity of Lille, 59000 Lille, FranceUniv. Lille, Inserm, CHU Lille, U1286–Infinite–Institute for Translational Research in Inflammation, 59000 Lille, FranceUniv. Lille, CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, US 41–UAR 2014–PLBS, 59000 Lille, FranceProteomics Platform Necker, Université Paris Cité–Structure Fédérative de Recherche Necker, INSERM US24/CNRS UAR3633, 75015 Paris, FranceProteomics Platform Necker, Université Paris Cité–Structure Fédérative de Recherche Necker, INSERM US24/CNRS UAR3633, 75015 Paris, FranceUniv. Lille, CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, U1019-UMR 9017-CIIL-Center for Infection and Immunity of Lille, 59000 Lille, FranceUniv. Lille, CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, U1019-UMR 9017-CIIL-Center for Infection and Immunity of Lille, 59000 Lille, FranceThe Protein Phosphatase type 1 catalytic subunit (PP1c) (PF3D7_1414400) operates in combination with various regulatory proteins to specifically direct and control its phosphatase activity. However, there is little information about this phosphatase and its regulators in the human malaria parasite, <i>Plasmodium falciparum</i>. To address this knowledge gap, we conducted a comprehensive investigation into the structural and functional characteristics of a conserved <i>Plasmodium</i>-specific regulator called Gametocyte EXported Protein 15, GEXP15 (PF3D7_1031600). Through in silico analysis, we identified three significant regions of interest in GEXP15: an N-terminal region housing a PP1-interacting RVxF motif, a conserved domain whose function is unknown, and a GYF-like domain that potentially facilitates specific protein–protein interactions. To further elucidate the role of GEXP15, we conducted in vitro interaction studies that demonstrated a direct interaction between GEXP15 and PP1 via the RVxF-binding motif. This interaction was found to enhance the phosphatase activity of PP1. Additionally, utilizing a transgenic GEXP15-tagged line and live microscopy, we observed high expression of GEXP15 in late asexual stages of the parasite, with localization predominantly in the nucleus. Immunoprecipitation assays followed by mass spectrometry analyses revealed the interaction of GEXP15 with ribosomal- and RNA-binding proteins. Furthermore, through pull-down analyses of recombinant functional domains of His-tagged GEXP15, we confirmed its binding to the ribosomal complex via the GYF domain. Collectively, our study sheds light on the PfGEXP15–PP1–ribosome interaction, which plays a crucial role in protein translation. These findings suggest that PfGEXP15 could serve as a potential target for the development of malaria drugs.https://www.mdpi.com/1422-0067/24/16/12647Protein Phosphatase 1PlasmodiummalariaGEXP15CD2BP2GYF domain |
| spellingShingle | Hala Mansour Alejandro Cabezas-Cruz Véronique Peucelle Amaury Farce Sophie Salomé-Desnoulez Ines Metatla Ida Chiara Guerrera Thomas Hollin Jamal Khalife Characterization of GEXP15 as a Potential Regulator of Protein Phosphatase 1 in <i>Plasmodium falciparum</i> International Journal of Molecular Sciences Protein Phosphatase 1 Plasmodium malaria GEXP15 CD2BP2 GYF domain |
| title | Characterization of GEXP15 as a Potential Regulator of Protein Phosphatase 1 in <i>Plasmodium falciparum</i> |
| title_full | Characterization of GEXP15 as a Potential Regulator of Protein Phosphatase 1 in <i>Plasmodium falciparum</i> |
| title_fullStr | Characterization of GEXP15 as a Potential Regulator of Protein Phosphatase 1 in <i>Plasmodium falciparum</i> |
| title_full_unstemmed | Characterization of GEXP15 as a Potential Regulator of Protein Phosphatase 1 in <i>Plasmodium falciparum</i> |
| title_short | Characterization of GEXP15 as a Potential Regulator of Protein Phosphatase 1 in <i>Plasmodium falciparum</i> |
| title_sort | characterization of gexp15 as a potential regulator of protein phosphatase 1 in i plasmodium falciparum i |
| topic | Protein Phosphatase 1 Plasmodium malaria GEXP15 CD2BP2 GYF domain |
| url | https://www.mdpi.com/1422-0067/24/16/12647 |
| work_keys_str_mv | AT halamansour characterizationofgexp15asapotentialregulatorofproteinphosphatase1iniplasmodiumfalciparumi AT alejandrocabezascruz characterizationofgexp15asapotentialregulatorofproteinphosphatase1iniplasmodiumfalciparumi AT veroniquepeucelle characterizationofgexp15asapotentialregulatorofproteinphosphatase1iniplasmodiumfalciparumi AT amauryfarce characterizationofgexp15asapotentialregulatorofproteinphosphatase1iniplasmodiumfalciparumi AT sophiesalomedesnoulez characterizationofgexp15asapotentialregulatorofproteinphosphatase1iniplasmodiumfalciparumi AT inesmetatla characterizationofgexp15asapotentialregulatorofproteinphosphatase1iniplasmodiumfalciparumi AT idachiaraguerrera characterizationofgexp15asapotentialregulatorofproteinphosphatase1iniplasmodiumfalciparumi AT thomashollin characterizationofgexp15asapotentialregulatorofproteinphosphatase1iniplasmodiumfalciparumi AT jamalkhalife characterizationofgexp15asapotentialregulatorofproteinphosphatase1iniplasmodiumfalciparumi |