N-terminal β-strand underpins biochemical specialization of an ATG8 isoform.
Autophagy-related protein 8 (ATG8) is a highly conserved ubiquitin-like protein that modulates autophagy pathways by binding autophagic membranes and a number of proteins, including cargo receptors and core autophagy components. Throughout plant evolution, ATG8 has expanded from a single protein in...
| Main Authors: | , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2019-07-01
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| Series: | PLoS Biology |
| Online Access: | https://doi.org/10.1371/journal.pbio.3000373 |
| _version_ | 1811191220490207232 |
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| author | Erin K Zess Cassandra Jensen Neftaly Cruz-Mireles Juan Carlos De la Concepcion Jan Sklenar Madlen Stephani Richard Imre Elisabeth Roitinger Richard Hughes Khaoula Belhaj Karl Mechtler Frank L H Menke Tolga Bozkurt Mark J Banfield Sophien Kamoun Abbas Maqbool Yasin F Dagdas |
| author_facet | Erin K Zess Cassandra Jensen Neftaly Cruz-Mireles Juan Carlos De la Concepcion Jan Sklenar Madlen Stephani Richard Imre Elisabeth Roitinger Richard Hughes Khaoula Belhaj Karl Mechtler Frank L H Menke Tolga Bozkurt Mark J Banfield Sophien Kamoun Abbas Maqbool Yasin F Dagdas |
| author_sort | Erin K Zess |
| collection | DOAJ |
| description | Autophagy-related protein 8 (ATG8) is a highly conserved ubiquitin-like protein that modulates autophagy pathways by binding autophagic membranes and a number of proteins, including cargo receptors and core autophagy components. Throughout plant evolution, ATG8 has expanded from a single protein in algae to multiple isoforms in higher plants. However, the degree to which ATG8 isoforms have functionally specialized to bind distinct proteins remains unclear. Here, we describe a comprehensive protein-protein interaction resource, obtained using in planta immunoprecipitation (IP) followed by mass spectrometry (MS), to define the potato ATG8 interactome. We discovered that ATG8 isoforms bind distinct sets of plant proteins with varying degrees of overlap. This prompted us to define the biochemical basis of ATG8 specialization by comparing two potato ATG8 isoforms using both in vivo protein interaction assays and in vitro quantitative binding affinity analyses. These experiments revealed that the N-terminal β-strand-and, in particular, a single amino acid polymorphism-underpins binding specificity to the substrate PexRD54 by shaping the hydrophobic pocket that accommodates this protein's ATG8-interacting motif (AIM). Additional proteomics experiments indicated that the N-terminal β-strand shapes the broader ATG8 interactor profiles, defining interaction specificity with about 80 plant proteins. Our findings are consistent with the view that ATG8 isoforms comprise a layer of specificity in the regulation of selective autophagy pathways in plants. |
| first_indexed | 2024-04-11T15:01:41Z |
| format | Article |
| id | doaj.art-6e8e206722e14333888edff117adeafb |
| institution | Directory Open Access Journal |
| issn | 1544-9173 1545-7885 |
| language | English |
| last_indexed | 2024-04-11T15:01:41Z |
| publishDate | 2019-07-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Biology |
| spelling | doaj.art-6e8e206722e14333888edff117adeafb2022-12-22T04:16:55ZengPublic Library of Science (PLoS)PLoS Biology1544-91731545-78852019-07-01177e300037310.1371/journal.pbio.3000373N-terminal β-strand underpins biochemical specialization of an ATG8 isoform.Erin K ZessCassandra JensenNeftaly Cruz-MirelesJuan Carlos De la ConcepcionJan SklenarMadlen StephaniRichard ImreElisabeth RoitingerRichard HughesKhaoula BelhajKarl MechtlerFrank L H MenkeTolga BozkurtMark J BanfieldSophien KamounAbbas MaqboolYasin F DagdasAutophagy-related protein 8 (ATG8) is a highly conserved ubiquitin-like protein that modulates autophagy pathways by binding autophagic membranes and a number of proteins, including cargo receptors and core autophagy components. Throughout plant evolution, ATG8 has expanded from a single protein in algae to multiple isoforms in higher plants. However, the degree to which ATG8 isoforms have functionally specialized to bind distinct proteins remains unclear. Here, we describe a comprehensive protein-protein interaction resource, obtained using in planta immunoprecipitation (IP) followed by mass spectrometry (MS), to define the potato ATG8 interactome. We discovered that ATG8 isoforms bind distinct sets of plant proteins with varying degrees of overlap. This prompted us to define the biochemical basis of ATG8 specialization by comparing two potato ATG8 isoforms using both in vivo protein interaction assays and in vitro quantitative binding affinity analyses. These experiments revealed that the N-terminal β-strand-and, in particular, a single amino acid polymorphism-underpins binding specificity to the substrate PexRD54 by shaping the hydrophobic pocket that accommodates this protein's ATG8-interacting motif (AIM). Additional proteomics experiments indicated that the N-terminal β-strand shapes the broader ATG8 interactor profiles, defining interaction specificity with about 80 plant proteins. Our findings are consistent with the view that ATG8 isoforms comprise a layer of specificity in the regulation of selective autophagy pathways in plants.https://doi.org/10.1371/journal.pbio.3000373 |
| spellingShingle | Erin K Zess Cassandra Jensen Neftaly Cruz-Mireles Juan Carlos De la Concepcion Jan Sklenar Madlen Stephani Richard Imre Elisabeth Roitinger Richard Hughes Khaoula Belhaj Karl Mechtler Frank L H Menke Tolga Bozkurt Mark J Banfield Sophien Kamoun Abbas Maqbool Yasin F Dagdas N-terminal β-strand underpins biochemical specialization of an ATG8 isoform. PLoS Biology |
| title | N-terminal β-strand underpins biochemical specialization of an ATG8 isoform. |
| title_full | N-terminal β-strand underpins biochemical specialization of an ATG8 isoform. |
| title_fullStr | N-terminal β-strand underpins biochemical specialization of an ATG8 isoform. |
| title_full_unstemmed | N-terminal β-strand underpins biochemical specialization of an ATG8 isoform. |
| title_short | N-terminal β-strand underpins biochemical specialization of an ATG8 isoform. |
| title_sort | n terminal β strand underpins biochemical specialization of an atg8 isoform |
| url | https://doi.org/10.1371/journal.pbio.3000373 |
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