Defective recognition of ATG8/LC3B by mutant SQSTM1/p62 implicates impairment of autophagy as a pathogenic mechanism in ALS-FTLD
Growing evidence implicates impairment of autophagy as a candidate pathogenic mechanism in the spectrum of neurodegenerative disorders which includes amyotrophic lateral sclerosis and frontotemporal lobar degeneration (ALS-FTLD). SQSTM1, which encodes the autophagy receptor SQSTM1/p62, is geneticall...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
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Taylor & Francis
2016
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| Online Access: | https://eprints.nottingham.ac.uk/31167/ |
| _version_ | 1848794140756672512 |
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| author | Goode, Alice Butler, Kevin Long, Jed Cavey, James Daniel, Scott Shaw, Barry Sollenberger, Jill Gell, Christopher Johansen, Terje Oldham, Neil J. Searle, Mark Layfield, Robert |
| author_facet | Goode, Alice Butler, Kevin Long, Jed Cavey, James Daniel, Scott Shaw, Barry Sollenberger, Jill Gell, Christopher Johansen, Terje Oldham, Neil J. Searle, Mark Layfield, Robert |
| author_sort | Goode, Alice |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | Growing evidence implicates impairment of autophagy as a candidate pathogenic mechanism in the spectrum of neurodegenerative disorders which includes amyotrophic lateral sclerosis and frontotemporal lobar degeneration (ALS-FTLD). SQSTM1, which encodes the autophagy receptor SQSTM1/p62, is genetically associated with ALS-FTLD, although to date autophagy-relevant functional defects in disease-associated variants have not been described. A key protein-protein interaction in autophagy is the recognition of lipid-anchored ATG8/LC3 within the phagophore membrane by SQSTM1, mediated through its LC3-interacting region (LIR), and notably some ALS-FTLD mutations map to this region. Here we show that although representing a conservative substitution and predicted to be benign, the ALS-associated L341V mutation of SQSTM1 is defective in recognition of LC3B. We place our observations on a firm quantitative footing by showing the L341V-mutant LIR is associated with a ~3-fold reduction in LC3B binding affinity and using protein NMR we rationalise the structural basis for the effect. This functional deficit is realised in motor neurone-like cells, with L341V mutant EGFP-mCherry-SQSTM1 less readily incorporated into acidic autophagic vesicles than wild-type. Our data supports a model in which the L341V mutation limits the critical step of SQSTM1 recruitment to the phagophore. The oligomeric nature of SQSTM1 which presents multiple LIRs to template growth of the phagophore potentially gives rise to avidity effects which amplify the relatively modest impact of any single mutation on LC3B binding. Over the lifetime of a neurone impaired autophagy could expose a vulnerability which ultimately tips the balance from cell survival towards cell death. |
| first_indexed | 2025-11-14T19:11:28Z |
| format | Article |
| id | nottingham-31167 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:11:28Z |
| publishDate | 2016 |
| publisher | Taylor & Francis |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-311672020-05-04T18:03:53Z https://eprints.nottingham.ac.uk/31167/ Defective recognition of ATG8/LC3B by mutant SQSTM1/p62 implicates impairment of autophagy as a pathogenic mechanism in ALS-FTLD Goode, Alice Butler, Kevin Long, Jed Cavey, James Daniel, Scott Shaw, Barry Sollenberger, Jill Gell, Christopher Johansen, Terje Oldham, Neil J. Searle, Mark Layfield, Robert Growing evidence implicates impairment of autophagy as a candidate pathogenic mechanism in the spectrum of neurodegenerative disorders which includes amyotrophic lateral sclerosis and frontotemporal lobar degeneration (ALS-FTLD). SQSTM1, which encodes the autophagy receptor SQSTM1/p62, is genetically associated with ALS-FTLD, although to date autophagy-relevant functional defects in disease-associated variants have not been described. A key protein-protein interaction in autophagy is the recognition of lipid-anchored ATG8/LC3 within the phagophore membrane by SQSTM1, mediated through its LC3-interacting region (LIR), and notably some ALS-FTLD mutations map to this region. Here we show that although representing a conservative substitution and predicted to be benign, the ALS-associated L341V mutation of SQSTM1 is defective in recognition of LC3B. We place our observations on a firm quantitative footing by showing the L341V-mutant LIR is associated with a ~3-fold reduction in LC3B binding affinity and using protein NMR we rationalise the structural basis for the effect. This functional deficit is realised in motor neurone-like cells, with L341V mutant EGFP-mCherry-SQSTM1 less readily incorporated into acidic autophagic vesicles than wild-type. Our data supports a model in which the L341V mutation limits the critical step of SQSTM1 recruitment to the phagophore. The oligomeric nature of SQSTM1 which presents multiple LIRs to template growth of the phagophore potentially gives rise to avidity effects which amplify the relatively modest impact of any single mutation on LC3B binding. Over the lifetime of a neurone impaired autophagy could expose a vulnerability which ultimately tips the balance from cell survival towards cell death. Taylor & Francis 2016-07-02 Article PeerReviewed Goode, Alice, Butler, Kevin, Long, Jed, Cavey, James, Daniel, Scott, Shaw, Barry, Sollenberger, Jill, Gell, Christopher, Johansen, Terje, Oldham, Neil J., Searle, Mark and Layfield, Robert (2016) Defective recognition of ATG8/LC3B by mutant SQSTM1/p62 implicates impairment of autophagy as a pathogenic mechanism in ALS-FTLD. Autophagy, 12 (7). ISSN 1554-8627 http://www.tandfonline.com/doi/full/10.1080/15548627.2016.1170257 doi:10.1080/15548627.2016.1170257 doi:10.1080/15548627.2016.1170257 |
| spellingShingle | Goode, Alice Butler, Kevin Long, Jed Cavey, James Daniel, Scott Shaw, Barry Sollenberger, Jill Gell, Christopher Johansen, Terje Oldham, Neil J. Searle, Mark Layfield, Robert Defective recognition of ATG8/LC3B by mutant SQSTM1/p62 implicates impairment of autophagy as a pathogenic mechanism in ALS-FTLD |
| title | Defective recognition of ATG8/LC3B by mutant SQSTM1/p62 implicates impairment of autophagy as a pathogenic mechanism in ALS-FTLD |
| title_full | Defective recognition of ATG8/LC3B by mutant SQSTM1/p62 implicates impairment of autophagy as a pathogenic mechanism in ALS-FTLD |
| title_fullStr | Defective recognition of ATG8/LC3B by mutant SQSTM1/p62 implicates impairment of autophagy as a pathogenic mechanism in ALS-FTLD |
| title_full_unstemmed | Defective recognition of ATG8/LC3B by mutant SQSTM1/p62 implicates impairment of autophagy as a pathogenic mechanism in ALS-FTLD |
| title_short | Defective recognition of ATG8/LC3B by mutant SQSTM1/p62 implicates impairment of autophagy as a pathogenic mechanism in ALS-FTLD |
| title_sort | defective recognition of atg8/lc3b by mutant sqstm1/p62 implicates impairment of autophagy as a pathogenic mechanism in als-ftld |
| url | https://eprints.nottingham.ac.uk/31167/ https://eprints.nottingham.ac.uk/31167/ https://eprints.nottingham.ac.uk/31167/ |