The inhibitory mechanism of Hal3 on the yeast Ppz1 phosphatase
The Ser/Thr protein phosphatase (PPase) Ppz1 is an enzyme related to the ubiquitous type-1 PPases (PP1c) but found only in fungi. It is regulated by an inhibitory subunit, Hal3, which binds to its catalytic domain. Overexpression of Ppz1 is highly toxic for yeast cells, so its de-regulation has been...
| Authors: | ; ; |
|---|---|
| Format: | Article |
| Publication date: | 2017 |
| Country: | España |
| Institution: | Universitat Autònoma de Barcelona |
| Repository: | Dipòsit Digital de Documents de la UAB |
| Language: | English |
| OAI Identifier: | oai:ddd.uab.cat:205909 |
| Online Access: | https://ddd.uab.cat/record/205909 https://dx.doi.org/urn:doi:10.1038/s41598-017-09360-5 |
| Access Level: | Open access |
| _version_ | 1878733045248294912 |
|---|---|
| author | Molero Merinero, Cristina |
| author2 | Casado Vázquez, Carlos Ariño Carmona, Joaquín|||0000-0002-6774-2987 |
| author2_role | author author |
| author_browse | Ariño Carmona, Joaquín|||0000-0002-6774-2987 Casado Vázquez, Carlos Molero Merinero, Cristina |
| author_facet | Molero Merinero, Cristina Casado Vázquez, Carlos Ariño Carmona, Joaquín|||0000-0002-6774-2987 |
| author_role | author |
| collection | Dipòsit Digital de Documents de la UAB |
| description | The Ser/Thr protein phosphatase (PPase) Ppz1 is an enzyme related to the ubiquitous type-1 PPases (PP1c) but found only in fungi. It is regulated by an inhibitory subunit, Hal3, which binds to its catalytic domain. Overexpression of Ppz1 is highly toxic for yeast cells, so its de-regulation has been proposed as a target for novel antifungal therapies. While modulation of PP1c by its many regulatory subunits has been extensively characterized, the manner by which Hal3 controls Ppz1 remains unknown. We have used error-prone PCR mutagenesis to construct a library of Ppz1 variants and developed a functional assay to identify mutations afecting the binding or/and the inhibitory capacity of Hal3. We have characterized diverse Ppz1 mutated versions in vivo and in vitro and found that, although they were clearly refractory to Hal3 inhibition, none of them exhibited signifcant reduction in Hal3 binding. Mapping the mutations strengthened the notion that Hal3 does not interact with Ppz1 through its RVxF-like motif (found in most PP1c regulators). In contrast, the most relevant mutations mapped to a conserved α-helix region used by mammalian Inhibitor-2 to regulate PP1c. Therefore, modulation of PP1c and Ppz1 by their subunits likely difers, but could share some structural features. |
| eu_rights_str_mv | openAccess |
| format | article |
| id | ES_4f00a22dfd650740f78aa7ce7b0785b0 |
| institution | Universitat Autònoma de Barcelona |
| instname_str | Universitat Autònoma de Barcelona |
| language | eng |
| network_acronym_str | ES |
| network_name_str | España |
| oai_identifier_str | oai:ddd.uab.cat:205909 |
| publishDate | 2017 |
| publishDateSort | 2017 |
| reponame_str | Dipòsit Digital de Documents de la UAB |
| spelling | The inhibitory mechanism of Hal3 on the yeast Ppz1 phosphatase A mutagenesis analysis Molero Merinero, Cristina Casado Vázquez, Carlos Ariño Carmona, Joaquín|||0000-0002-6774-2987 The Ser/Thr protein phosphatase (PPase) Ppz1 is an enzyme related to the ubiquitous type-1 PPases (PP1c) but found only in fungi. It is regulated by an inhibitory subunit, Hal3, which binds to its catalytic domain. Overexpression of Ppz1 is highly toxic for yeast cells, so its de-regulation has been proposed as a target for novel antifungal therapies. While modulation of PP1c by its many regulatory subunits has been extensively characterized, the manner by which Hal3 controls Ppz1 remains unknown. We have used error-prone PCR mutagenesis to construct a library of Ppz1 variants and developed a functional assay to identify mutations afecting the binding or/and the inhibitory capacity of Hal3. We have characterized diverse Ppz1 mutated versions in vivo and in vitro and found that, although they were clearly refractory to Hal3 inhibition, none of them exhibited signifcant reduction in Hal3 binding. Mapping the mutations strengthened the notion that Hal3 does not interact with Ppz1 through its RVxF-like motif (found in most PP1c regulators). In contrast, the most relevant mutations mapped to a conserved α-helix region used by mammalian Inhibitor-2 to regulate PP1c. Therefore, modulation of PP1c and Ppz1 by their subunits likely difers, but could share some structural features. https://ddd.uab.cat/record/205909 https://dx.doi.org/urn:doi:10.1038/s41598-017-09360-5 |
| spellingShingle | The inhibitory mechanism of Hal3 on the yeast Ppz1 phosphatase Molero Merinero, Cristina |
| title | The inhibitory mechanism of Hal3 on the yeast Ppz1 phosphatase |
| title_full | The inhibitory mechanism of Hal3 on the yeast Ppz1 phosphatase |
| title_fullStr | The inhibitory mechanism of Hal3 on the yeast Ppz1 phosphatase |
| title_full_unstemmed | The inhibitory mechanism of Hal3 on the yeast Ppz1 phosphatase |
| title_short | The inhibitory mechanism of Hal3 on the yeast Ppz1 phosphatase |
| title_sort | The inhibitory mechanism of Hal3 on the yeast Ppz1 phosphatase |
| url | https://ddd.uab.cat/record/205909 https://dx.doi.org/urn:doi:10.1038/s41598-017-09360-5 |