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Theileria parasites secrete a prolyl isomerase to maintain host leukocyte transformation.

Marsolier J, Perichon M, DeBarry JD, Villoutreix BO, Chluba J, Lopez T, Garrido C, Zhou XZ, Lu KP, Fritsch L, Ait-Si-Ali S, Mhadhbi M, Medjkane S, Weitzman JB - Nature (2015)

Bottom Line: Here we show that TaPIN1 is a bona fide prolyl isomerase and that it interacts with the host ubiquitin ligase FBW7, leading to its degradation and subsequent stabilization of c-JUN, which promotes transformation.We performed in vitro and in silico analysis and in vivo zebrafish xenograft experiments to demonstrate that TaPIN1 is directly inhibited by the anti-parasite drug buparvaquone (and other known PIN1 inhibitors) and is mutated in a drug-resistant strain.Prolyl isomerization is thus a conserved mechanism that is important in cancer and is used by Theileria parasites to manipulate host oncogenic signalling.

View Article: PubMed Central - PubMed

Affiliation: Université Paris Diderot, Sorbonne Paris Cité, Epigenetics and Cell Fate, UMR 7216 CNRS, 75013 Paris, France.

ABSTRACT
Infectious agents develop intricate mechanisms to interact with host cell pathways and hijack their genetic and epigenetic machinery to change host cell phenotypic states. Among the Apicomplexa phylum of obligate intracellular parasites, which cause veterinary and human diseases, Theileria is the only genus that transforms its mammalian host cells. Theileria infection of bovine leukocytes induces proliferative and invasive phenotypes associated with activated signalling pathways, notably JNK and AP-1 (ref. 2). The transformed phenotypes are reversed by treatment with the theilericidal drug buparvaquone. We used comparative genomics to identify a homologue of the peptidyl-prolyl isomerase PIN1 in T. annulata (TaPIN1) that is secreted into the host cell and modulates oncogenic signalling pathways. Here we show that TaPIN1 is a bona fide prolyl isomerase and that it interacts with the host ubiquitin ligase FBW7, leading to its degradation and subsequent stabilization of c-JUN, which promotes transformation. We performed in vitro and in silico analysis and in vivo zebrafish xenograft experiments to demonstrate that TaPIN1 is directly inhibited by the anti-parasite drug buparvaquone (and other known PIN1 inhibitors) and is mutated in a drug-resistant strain. Prolyl isomerization is thus a conserved mechanism that is important in cancer and is used by Theileria parasites to manipulate host oncogenic signalling.

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The PPIase domain of Pin1 is well conserveda. Sequence alignment of pin1 genes in H. sapiens, A. thaliana, T. brucei and T. annulata revealed the presence or the absence of a conserved WW domain. Percentages of identity are indicated. The magenta box indicates the predicted signal peptide of TaPin1.b. Homology models for TaPin1 WT and Mutant A53P based on sequence identity/similarity with hPin1. The mutation Ala>Pro in TaPin1 appears to induce a conformational change within and nearby the catalytic loop.c. The Buparvaquone molecule can be docked in the active site of hPin1 or in the active site of WT TaPin1 predicted structure. Here, the second lowest docked energy pose is shown (the best predicted energy pose is shown Fig. 3a). These two predicted binding poses are fully consistent with the different binding modes of some inhibitors co-crystallized with hPin1. Yet, considering computations shown below in Extended Data Fig. 6e, we suggest that the most likely pose for Buparvaquone corresponds to the pose 1 reported in Fig 3a. However, independently of the selected poses, we expect that Buparvaquone would not fit well in the catalytic pocket due to structural changes induced by the A53P mutation.d. 3D structure of the experimental hPin1 structure and the predicted TaPin1 WT, A53P mutant, alongside Trypanosome TbPin1, Arabidopsis Pin1At models (ribbon diagram). The 3D structures are well conserved among these proteins with some differences, for instance, in the catalytic loops. The TaPin1, TbPin1 and PinAt enzymes lack the WW domain present in hPin1 yet the overall fold in the catalytic area is well-conserved suggesting that accurate homology models for TaPin1 can be built using the approach used in the present study.e. The experimental structure of hPin1 is represented as a solid surface with a view down the active site, showing a small co-crystallized ligand next to the catalytic site residue Cys113. In the same orientation, small chemical fragments are predicted to bind in the catalytic site region with FTmap. Using this information, we propose that the most likely binding pose for Buparvaquone is the one shown in Fig. 3.
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Figure 10: The PPIase domain of Pin1 is well conserveda. Sequence alignment of pin1 genes in H. sapiens, A. thaliana, T. brucei and T. annulata revealed the presence or the absence of a conserved WW domain. Percentages of identity are indicated. The magenta box indicates the predicted signal peptide of TaPin1.b. Homology models for TaPin1 WT and Mutant A53P based on sequence identity/similarity with hPin1. The mutation Ala>Pro in TaPin1 appears to induce a conformational change within and nearby the catalytic loop.c. The Buparvaquone molecule can be docked in the active site of hPin1 or in the active site of WT TaPin1 predicted structure. Here, the second lowest docked energy pose is shown (the best predicted energy pose is shown Fig. 3a). These two predicted binding poses are fully consistent with the different binding modes of some inhibitors co-crystallized with hPin1. Yet, considering computations shown below in Extended Data Fig. 6e, we suggest that the most likely pose for Buparvaquone corresponds to the pose 1 reported in Fig 3a. However, independently of the selected poses, we expect that Buparvaquone would not fit well in the catalytic pocket due to structural changes induced by the A53P mutation.d. 3D structure of the experimental hPin1 structure and the predicted TaPin1 WT, A53P mutant, alongside Trypanosome TbPin1, Arabidopsis Pin1At models (ribbon diagram). The 3D structures are well conserved among these proteins with some differences, for instance, in the catalytic loops. The TaPin1, TbPin1 and PinAt enzymes lack the WW domain present in hPin1 yet the overall fold in the catalytic area is well-conserved suggesting that accurate homology models for TaPin1 can be built using the approach used in the present study.e. The experimental structure of hPin1 is represented as a solid surface with a view down the active site, showing a small co-crystallized ligand next to the catalytic site residue Cys113. In the same orientation, small chemical fragments are predicted to bind in the catalytic site region with FTmap. Using this information, we propose that the most likely binding pose for Buparvaquone is the one shown in Fig. 3.

Mentions: In a search for potential inhibitors, we noted that the chemical structure of Buparvaquone is similar to Juglone, a well-characterized inhibitor of mammalian Pin113. The TaPin1 sequence exhibits over 47% identity with hPin1 in the PPIase domain (Extended Data Fig. 6a). Our homology models of TaPin1 protein based on published hPin1 experimental data suggest a similar structure with a conserved catalytic pocket (Fig. 3a, Extended Data Fig. 6b). Notably, several Pin1 homologues also lack the WW domain, including Arabidopsis thaliana Pin1At18-20, MdPin1 in Malus domestica and the parasite Trypanosoma brucei TbPin1 homologue20-22, and the predicted TaPin1 model closely resembles these structures (Extended Data Fig. 6d). We investigated the hPin1 experimental structure and the TaPin1 predicted model with the binding pocket and hot-spot detection algorithm FTMap, using the server FTFlex. Notably, we found key hot-spot regions in the catalytic site area, matching the substrate binding region of hPin1 (Extended Data Fig. 6). Juglone and Buparvaquone molecules could be docked into the active site of both TaPin1 and hPin1 by in silico analysis (Fig. 3a, Extended Data Fig. 6c).


Theileria parasites secrete a prolyl isomerase to maintain host leukocyte transformation.

Marsolier J, Perichon M, DeBarry JD, Villoutreix BO, Chluba J, Lopez T, Garrido C, Zhou XZ, Lu KP, Fritsch L, Ait-Si-Ali S, Mhadhbi M, Medjkane S, Weitzman JB - Nature (2015)

The PPIase domain of Pin1 is well conserveda. Sequence alignment of pin1 genes in H. sapiens, A. thaliana, T. brucei and T. annulata revealed the presence or the absence of a conserved WW domain. Percentages of identity are indicated. The magenta box indicates the predicted signal peptide of TaPin1.b. Homology models for TaPin1 WT and Mutant A53P based on sequence identity/similarity with hPin1. The mutation Ala>Pro in TaPin1 appears to induce a conformational change within and nearby the catalytic loop.c. The Buparvaquone molecule can be docked in the active site of hPin1 or in the active site of WT TaPin1 predicted structure. Here, the second lowest docked energy pose is shown (the best predicted energy pose is shown Fig. 3a). These two predicted binding poses are fully consistent with the different binding modes of some inhibitors co-crystallized with hPin1. Yet, considering computations shown below in Extended Data Fig. 6e, we suggest that the most likely pose for Buparvaquone corresponds to the pose 1 reported in Fig 3a. However, independently of the selected poses, we expect that Buparvaquone would not fit well in the catalytic pocket due to structural changes induced by the A53P mutation.d. 3D structure of the experimental hPin1 structure and the predicted TaPin1 WT, A53P mutant, alongside Trypanosome TbPin1, Arabidopsis Pin1At models (ribbon diagram). The 3D structures are well conserved among these proteins with some differences, for instance, in the catalytic loops. The TaPin1, TbPin1 and PinAt enzymes lack the WW domain present in hPin1 yet the overall fold in the catalytic area is well-conserved suggesting that accurate homology models for TaPin1 can be built using the approach used in the present study.e. The experimental structure of hPin1 is represented as a solid surface with a view down the active site, showing a small co-crystallized ligand next to the catalytic site residue Cys113. In the same orientation, small chemical fragments are predicted to bind in the catalytic site region with FTmap. Using this information, we propose that the most likely binding pose for Buparvaquone is the one shown in Fig. 3.
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Figure 10: The PPIase domain of Pin1 is well conserveda. Sequence alignment of pin1 genes in H. sapiens, A. thaliana, T. brucei and T. annulata revealed the presence or the absence of a conserved WW domain. Percentages of identity are indicated. The magenta box indicates the predicted signal peptide of TaPin1.b. Homology models for TaPin1 WT and Mutant A53P based on sequence identity/similarity with hPin1. The mutation Ala>Pro in TaPin1 appears to induce a conformational change within and nearby the catalytic loop.c. The Buparvaquone molecule can be docked in the active site of hPin1 or in the active site of WT TaPin1 predicted structure. Here, the second lowest docked energy pose is shown (the best predicted energy pose is shown Fig. 3a). These two predicted binding poses are fully consistent with the different binding modes of some inhibitors co-crystallized with hPin1. Yet, considering computations shown below in Extended Data Fig. 6e, we suggest that the most likely pose for Buparvaquone corresponds to the pose 1 reported in Fig 3a. However, independently of the selected poses, we expect that Buparvaquone would not fit well in the catalytic pocket due to structural changes induced by the A53P mutation.d. 3D structure of the experimental hPin1 structure and the predicted TaPin1 WT, A53P mutant, alongside Trypanosome TbPin1, Arabidopsis Pin1At models (ribbon diagram). The 3D structures are well conserved among these proteins with some differences, for instance, in the catalytic loops. The TaPin1, TbPin1 and PinAt enzymes lack the WW domain present in hPin1 yet the overall fold in the catalytic area is well-conserved suggesting that accurate homology models for TaPin1 can be built using the approach used in the present study.e. The experimental structure of hPin1 is represented as a solid surface with a view down the active site, showing a small co-crystallized ligand next to the catalytic site residue Cys113. In the same orientation, small chemical fragments are predicted to bind in the catalytic site region with FTmap. Using this information, we propose that the most likely binding pose for Buparvaquone is the one shown in Fig. 3.
Mentions: In a search for potential inhibitors, we noted that the chemical structure of Buparvaquone is similar to Juglone, a well-characterized inhibitor of mammalian Pin113. The TaPin1 sequence exhibits over 47% identity with hPin1 in the PPIase domain (Extended Data Fig. 6a). Our homology models of TaPin1 protein based on published hPin1 experimental data suggest a similar structure with a conserved catalytic pocket (Fig. 3a, Extended Data Fig. 6b). Notably, several Pin1 homologues also lack the WW domain, including Arabidopsis thaliana Pin1At18-20, MdPin1 in Malus domestica and the parasite Trypanosoma brucei TbPin1 homologue20-22, and the predicted TaPin1 model closely resembles these structures (Extended Data Fig. 6d). We investigated the hPin1 experimental structure and the TaPin1 predicted model with the binding pocket and hot-spot detection algorithm FTMap, using the server FTFlex. Notably, we found key hot-spot regions in the catalytic site area, matching the substrate binding region of hPin1 (Extended Data Fig. 6). Juglone and Buparvaquone molecules could be docked into the active site of both TaPin1 and hPin1 by in silico analysis (Fig. 3a, Extended Data Fig. 6c).

Bottom Line: Here we show that TaPIN1 is a bona fide prolyl isomerase and that it interacts with the host ubiquitin ligase FBW7, leading to its degradation and subsequent stabilization of c-JUN, which promotes transformation.We performed in vitro and in silico analysis and in vivo zebrafish xenograft experiments to demonstrate that TaPIN1 is directly inhibited by the anti-parasite drug buparvaquone (and other known PIN1 inhibitors) and is mutated in a drug-resistant strain.Prolyl isomerization is thus a conserved mechanism that is important in cancer and is used by Theileria parasites to manipulate host oncogenic signalling.

View Article: PubMed Central - PubMed

Affiliation: Université Paris Diderot, Sorbonne Paris Cité, Epigenetics and Cell Fate, UMR 7216 CNRS, 75013 Paris, France.

ABSTRACT
Infectious agents develop intricate mechanisms to interact with host cell pathways and hijack their genetic and epigenetic machinery to change host cell phenotypic states. Among the Apicomplexa phylum of obligate intracellular parasites, which cause veterinary and human diseases, Theileria is the only genus that transforms its mammalian host cells. Theileria infection of bovine leukocytes induces proliferative and invasive phenotypes associated with activated signalling pathways, notably JNK and AP-1 (ref. 2). The transformed phenotypes are reversed by treatment with the theilericidal drug buparvaquone. We used comparative genomics to identify a homologue of the peptidyl-prolyl isomerase PIN1 in T. annulata (TaPIN1) that is secreted into the host cell and modulates oncogenic signalling pathways. Here we show that TaPIN1 is a bona fide prolyl isomerase and that it interacts with the host ubiquitin ligase FBW7, leading to its degradation and subsequent stabilization of c-JUN, which promotes transformation. We performed in vitro and in silico analysis and in vivo zebrafish xenograft experiments to demonstrate that TaPIN1 is directly inhibited by the anti-parasite drug buparvaquone (and other known PIN1 inhibitors) and is mutated in a drug-resistant strain. Prolyl isomerization is thus a conserved mechanism that is important in cancer and is used by Theileria parasites to manipulate host oncogenic signalling.

Show MeSH
Related in: MedlinePlus