Cloning and functional complementation of ten Schistosoma mansoni phosphodiesterases expressed in the mammalian host stages
Only a single drug against schistosomiasis is currently available and new drug development is urgently required but very few drug targets have been validated and characterised. However, regulatory systems including cyclic nucleotide metabolism are emerging as primary candidates for drug discovery. Here, we report the cloning of ten cyclic nucleotide phosphodiesterase (PDE) genes of S. mansoni, out of a total of 11 identified in its genome. We classify these PDEs by homology to human PDEs. Male worms displayed higher expression levels for all PDEs, in mature and juvenile worms, and schistosomula. Several functional complementation approaches were used to characterise these genes. We constructed a Trypanosoma brucei cell line in which expression of a cAMP-degrading PDE complements the deletion of TbrPDEB1/B2. Inhibitor screens of these cells expressing only either SmPDE4A, TbrPDEB1 or TbrPDEB2, identified highly potent inhibitors of the S. mansoni enzyme that elevated the cellular cAMP concentration. We further expressed most of the cloned SmPDEs in two pde1Δ/pde2Δ strains of Saccharomyces cerevisiae and some also in a specialised strain of Schizosacharomyces pombe. Five PDEs, SmPDE1, SmPDE4A, SmPDE8, SmPDE9A and SmPDE11 successfully complemented the S. cerevisiae strains, and SmPDE7var also complemented to a lesser degree, in liquid culture. SmPDE4A, SmPDE8 and SmPDE11 were further assessed in S. pombe for hydrolysis of cAMP and cGMP; SmPDE11 displayed considerable preferrence for cGMP over cAMP. These results and tools enable the pursuit of a rigorous drug discovery program based on inhibitors of S. mansoni PDEs.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2020 |
|---|---|
| Erschienen: |
2020 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:14 |
|---|---|
| Enthalten in: |
PLoS neglected tropical diseases - 14(2020), 7 vom: 14. Juli, Seite e0008447 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Munday, Jane C [VerfasserIn] |
|---|
| Links: |
|---|
| Themen: |
EC 3.1.4.- |
|---|
| Anmerkungen: |
Date Completed 11.09.2020 Date Revised 29.03.2024 published: Electronic-eCollection Citation Status MEDLINE |
|---|
| doi: |
10.1371/journal.pntd.0008447 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM313059020 |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM313059020 | ||
| 003 | DE-627 | ||
| 005 | 20240329233855.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231225s2020 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1371/journal.pntd.0008447 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1354.xml |
| 035 | |a (DE-627)NLM313059020 | ||
| 035 | |a (NLM)32730343 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Munday, Jane C |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Cloning and functional complementation of ten Schistosoma mansoni phosphodiesterases expressed in the mammalian host stages |
| 264 | 1 | |c 2020 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 11.09.2020 | ||
| 500 | |a Date Revised 29.03.2024 | ||
| 500 | |a published: Electronic-eCollection | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Only a single drug against schistosomiasis is currently available and new drug development is urgently required but very few drug targets have been validated and characterised. However, regulatory systems including cyclic nucleotide metabolism are emerging as primary candidates for drug discovery. Here, we report the cloning of ten cyclic nucleotide phosphodiesterase (PDE) genes of S. mansoni, out of a total of 11 identified in its genome. We classify these PDEs by homology to human PDEs. Male worms displayed higher expression levels for all PDEs, in mature and juvenile worms, and schistosomula. Several functional complementation approaches were used to characterise these genes. We constructed a Trypanosoma brucei cell line in which expression of a cAMP-degrading PDE complements the deletion of TbrPDEB1/B2. Inhibitor screens of these cells expressing only either SmPDE4A, TbrPDEB1 or TbrPDEB2, identified highly potent inhibitors of the S. mansoni enzyme that elevated the cellular cAMP concentration. We further expressed most of the cloned SmPDEs in two pde1Δ/pde2Δ strains of Saccharomyces cerevisiae and some also in a specialised strain of Schizosacharomyces pombe. Five PDEs, SmPDE1, SmPDE4A, SmPDE8, SmPDE9A and SmPDE11 successfully complemented the S. cerevisiae strains, and SmPDE7var also complemented to a lesser degree, in liquid culture. SmPDE4A, SmPDE8 and SmPDE11 were further assessed in S. pombe for hydrolysis of cAMP and cGMP; SmPDE11 displayed considerable preferrence for cGMP over cAMP. These results and tools enable the pursuit of a rigorous drug discovery program based on inhibitors of S. mansoni PDEs | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 7 | |a Helminth Proteins |2 NLM | |
| 650 | 7 | |a Phosphoric Diester Hydrolases |2 NLM | |
| 650 | 7 | |a EC 3.1.4.- |2 NLM | |
| 700 | 1 | |a Kunz, Stefan |e verfasserin |4 aut | |
| 700 | 1 | |a Kalejaiye, Titilola D |e verfasserin |4 aut | |
| 700 | 1 | |a Siderius, Marco |e verfasserin |4 aut | |
| 700 | 1 | |a Schroeder, Susanne |e verfasserin |4 aut | |
| 700 | 1 | |a Paape, Daniel |e verfasserin |4 aut | |
| 700 | 1 | |a Alghamdi, Ali H |e verfasserin |4 aut | |
| 700 | 1 | |a Abbasi, Zainab |e verfasserin |4 aut | |
| 700 | 1 | |a Huang, Sheng Xiang |e verfasserin |4 aut | |
| 700 | 1 | |a Donachie, Anne-Marie |e verfasserin |4 aut | |
| 700 | 1 | |a William, Samia |e verfasserin |4 aut | |
| 700 | 1 | |a Sabra, Abdel Nasser |e verfasserin |4 aut | |
| 700 | 1 | |a Sterk, Geert Jan |e verfasserin |4 aut | |
| 700 | 1 | |a Botros, Sanaa S |e verfasserin |4 aut | |
| 700 | 1 | |a Brown, David G |e verfasserin |4 aut | |
| 700 | 1 | |a Hoffman, Charles S |e verfasserin |4 aut | |
| 700 | 1 | |a Leurs, Rob |e verfasserin |4 aut | |
| 700 | 1 | |a de Koning, Harry P |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t PLoS neglected tropical diseases |d 2007 |g 14(2020), 7 vom: 14. Juli, Seite e0008447 |w (DE-627)NLM17489287X |x 1935-2735 |7 nnns |
| 773 | 1 | 8 | |g volume:14 |g year:2020 |g number:7 |g day:14 |g month:07 |g pages:e0008447 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1371/journal.pntd.0008447 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 14 |j 2020 |e 7 |b 14 |c 07 |h e0008447 | ||