Genetic and Pharmacological Inhibition of Autophagy Increases the Monoubiquitination of Non-Photosynthetic Phospho<i>enol</i>pyruvate Carboxylase
Phospho<i>enol</i>pyruvate carboxylase (PEPC) is an enzyme with key roles in carbon and nitrogen metabolisms. The mechanisms that control enzyme stability and turnover are not well known. This paper investigates the degradation of PEPC via selective autophagy, including the role of the m...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-12-01
|
| Series: | Plants |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2223-7747/10/1/12 |
| _version_ | 1797543673593856000 |
|---|---|
| author | Guillermo Baena Ana B. Feria Luis Hernández-Huertas Jacinto Gandullo Cristina Echevarría José A. Monreal Sofía García-Mauriño |
| author_facet | Guillermo Baena Ana B. Feria Luis Hernández-Huertas Jacinto Gandullo Cristina Echevarría José A. Monreal Sofía García-Mauriño |
| author_sort | Guillermo Baena |
| collection | DOAJ |
| description | Phospho<i>enol</i>pyruvate carboxylase (PEPC) is an enzyme with key roles in carbon and nitrogen metabolisms. The mechanisms that control enzyme stability and turnover are not well known. This paper investigates the degradation of PEPC via selective autophagy, including the role of the monoubiquitination of the enzyme in this process. In Arabidopsis, the genetic inhibition of autophagy increases the amount of monoubiquitinated PEPC in the <i>atg2</i>, <i>atg5</i>, and <i>atg18a</i> lines. The same is observed in <i>nbr1</i>, which is deficient in a protein that recruits monoubiquitinated substrates for selective autophagy. In cultured tobacco cells, the chemical inhibition of the degradation of autophagic substrates increases the quantity of PEPC proteins. When the formation of the autophagosome is blocked with 3-methyladenine (3-MA), monoubiquitinated PEPC accumulates as a result. Finally, pull-down experiments with a truncated version of NBR1 demonstrate the recovery of intact and/or fragmented PEPC in Arabidopsis leaves and roots, as well as cultured tobacco cells. Taken together, the results show that a fraction of PEPC is cleaved via selective autophagy and that the monoubiquitination of the enzyme has a role in its recruitment towards this pathway. Although autophagy seems to be a minor pathway, the results presented here increase the knowledge about the role of monoubiquitination and the regulation of PEPC degradation. |
| first_indexed | 2024-03-10T13:49:00Z |
| format | Article |
| id | doaj.art-00e5bd7ca7954482b8f2b16c71ef0cd9 |
| institution | Directory Open Access Journal |
| issn | 2223-7747 |
| language | English |
| last_indexed | 2024-03-10T13:49:00Z |
| publishDate | 2020-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Plants |
| spelling | doaj.art-00e5bd7ca7954482b8f2b16c71ef0cd92023-11-21T02:19:05ZengMDPI AGPlants2223-77472020-12-011011210.3390/plants10010012Genetic and Pharmacological Inhibition of Autophagy Increases the Monoubiquitination of Non-Photosynthetic Phospho<i>enol</i>pyruvate CarboxylaseGuillermo Baena0Ana B. Feria1Luis Hernández-Huertas2Jacinto Gandullo3Cristina Echevarría4José A. Monreal5Sofía García-Mauriño6Departamento de Biología Vegetal y Ecología, Facultad de Biología, Universidad de Sevilla, Avenida Reina Mercedes nº 6, 41012 Seville, SpainDepartamento de Biología Vegetal y Ecología, Facultad de Biología, Universidad de Sevilla, Avenida Reina Mercedes nº 6, 41012 Seville, SpainDepartamento de Biología Vegetal y Ecología, Facultad de Biología, Universidad de Sevilla, Avenida Reina Mercedes nº 6, 41012 Seville, SpainDepartamento de Biología Vegetal y Ecología, Facultad de Biología, Universidad de Sevilla, Avenida Reina Mercedes nº 6, 41012 Seville, SpainDepartamento de Biología Vegetal y Ecología, Facultad de Biología, Universidad de Sevilla, Avenida Reina Mercedes nº 6, 41012 Seville, SpainDepartamento de Biología Vegetal y Ecología, Facultad de Biología, Universidad de Sevilla, Avenida Reina Mercedes nº 6, 41012 Seville, SpainDepartamento de Biología Vegetal y Ecología, Facultad de Biología, Universidad de Sevilla, Avenida Reina Mercedes nº 6, 41012 Seville, SpainPhospho<i>enol</i>pyruvate carboxylase (PEPC) is an enzyme with key roles in carbon and nitrogen metabolisms. The mechanisms that control enzyme stability and turnover are not well known. This paper investigates the degradation of PEPC via selective autophagy, including the role of the monoubiquitination of the enzyme in this process. In Arabidopsis, the genetic inhibition of autophagy increases the amount of monoubiquitinated PEPC in the <i>atg2</i>, <i>atg5</i>, and <i>atg18a</i> lines. The same is observed in <i>nbr1</i>, which is deficient in a protein that recruits monoubiquitinated substrates for selective autophagy. In cultured tobacco cells, the chemical inhibition of the degradation of autophagic substrates increases the quantity of PEPC proteins. When the formation of the autophagosome is blocked with 3-methyladenine (3-MA), monoubiquitinated PEPC accumulates as a result. Finally, pull-down experiments with a truncated version of NBR1 demonstrate the recovery of intact and/or fragmented PEPC in Arabidopsis leaves and roots, as well as cultured tobacco cells. Taken together, the results show that a fraction of PEPC is cleaved via selective autophagy and that the monoubiquitination of the enzyme has a role in its recruitment towards this pathway. Although autophagy seems to be a minor pathway, the results presented here increase the knowledge about the role of monoubiquitination and the regulation of PEPC degradation.https://www.mdpi.com/2223-7747/10/1/12autophagy<i>Arabidopsis thaliana</i><i>Nicotiana benthamiana</i>phospho<i>enol</i>pyruvate carboxylaseubiquitin |
| spellingShingle | Guillermo Baena Ana B. Feria Luis Hernández-Huertas Jacinto Gandullo Cristina Echevarría José A. Monreal Sofía García-Mauriño Genetic and Pharmacological Inhibition of Autophagy Increases the Monoubiquitination of Non-Photosynthetic Phospho<i>enol</i>pyruvate Carboxylase Plants autophagy <i>Arabidopsis thaliana</i> <i>Nicotiana benthamiana</i> phospho<i>enol</i>pyruvate carboxylase ubiquitin |
| title | Genetic and Pharmacological Inhibition of Autophagy Increases the Monoubiquitination of Non-Photosynthetic Phospho<i>enol</i>pyruvate Carboxylase |
| title_full | Genetic and Pharmacological Inhibition of Autophagy Increases the Monoubiquitination of Non-Photosynthetic Phospho<i>enol</i>pyruvate Carboxylase |
| title_fullStr | Genetic and Pharmacological Inhibition of Autophagy Increases the Monoubiquitination of Non-Photosynthetic Phospho<i>enol</i>pyruvate Carboxylase |
| title_full_unstemmed | Genetic and Pharmacological Inhibition of Autophagy Increases the Monoubiquitination of Non-Photosynthetic Phospho<i>enol</i>pyruvate Carboxylase |
| title_short | Genetic and Pharmacological Inhibition of Autophagy Increases the Monoubiquitination of Non-Photosynthetic Phospho<i>enol</i>pyruvate Carboxylase |
| title_sort | genetic and pharmacological inhibition of autophagy increases the monoubiquitination of non photosynthetic phospho i enol i pyruvate carboxylase |
| topic | autophagy <i>Arabidopsis thaliana</i> <i>Nicotiana benthamiana</i> phospho<i>enol</i>pyruvate carboxylase ubiquitin |
| url | https://www.mdpi.com/2223-7747/10/1/12 |
| work_keys_str_mv | AT guillermobaena geneticandpharmacologicalinhibitionofautophagyincreasesthemonoubiquitinationofnonphotosyntheticphosphoienolipyruvatecarboxylase AT anabferia geneticandpharmacologicalinhibitionofautophagyincreasesthemonoubiquitinationofnonphotosyntheticphosphoienolipyruvatecarboxylase AT luishernandezhuertas geneticandpharmacologicalinhibitionofautophagyincreasesthemonoubiquitinationofnonphotosyntheticphosphoienolipyruvatecarboxylase AT jacintogandullo geneticandpharmacologicalinhibitionofautophagyincreasesthemonoubiquitinationofnonphotosyntheticphosphoienolipyruvatecarboxylase AT cristinaechevarria geneticandpharmacologicalinhibitionofautophagyincreasesthemonoubiquitinationofnonphotosyntheticphosphoienolipyruvatecarboxylase AT joseamonreal geneticandpharmacologicalinhibitionofautophagyincreasesthemonoubiquitinationofnonphotosyntheticphosphoienolipyruvatecarboxylase AT sofiagarciamaurino geneticandpharmacologicalinhibitionofautophagyincreasesthemonoubiquitinationofnonphotosyntheticphosphoienolipyruvatecarboxylase |