γ-COPI mediates the retention of kAE1 G701D protein in Golgi apparatus – A mechanistic explanation of distal renal tubular acidosis associated with the G701D mutation
© 2017 The Author(s); published by Portland Press Limited on behalf of the Biochemical Society. Mutations of the solute carrier family 4 member 1 (SLC4A1) gene encoding kidney anion (chloride/bicarbonate ion) exchanger 1 (kAE1) can cause genetic distal renal tubular acidosis (dRTA). Different SLC4A1...
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th-mahidol.418332019-03-14T15:02:50Z γ-COPI mediates the retention of kAE1 G701D protein in Golgi apparatus – A mechanistic explanation of distal renal tubular acidosis associated with the G701D mutation Natapol Duangtum Mutita Junking Suratchanee Phadngam Nunghathai Sawasdee Andrea Castiglioni Komgrid Charngkaew Thawornchai Limjindaporn Ciro Isidoro Pa Thai Yenchitsomanus Mahidol University Università degli Studi del Piemonte Orientale Amedeo Avogadro, Novara Biochemistry, Genetics and Molecular Biology © 2017 The Author(s); published by Portland Press Limited on behalf of the Biochemical Society. Mutations of the solute carrier family 4 member 1 (SLC4A1) gene encoding kidney anion (chloride/bicarbonate ion) exchanger 1 (kAE1) can cause genetic distal renal tubular acidosis (dRTA). Different SLC4A1 mutations give rise to mutant kAE1 proteins with distinct defects in protein trafficking. The mutant kAE1 protein may be retained in endoplasmic reticulum (ER) or Golgi apparatus, or mis-targeted to the apical membrane, failing to display its function at the baso-lateral membrane. The ER-retained mutant kAE1 interacts with calnexin chaperone protein; disruption of this interaction permits the mutant kAE1 to reach the cell surface and display anion exchange activity. However, the mechanism of Golgi retention of mutant kAE1 G701D protein, which is otherwise functional, is still unclear. In the present study, we show that Golgi retention of kAE1 G701D is due to a stable interaction with the Golgi-resident protein, coat protein complex I (COPI), that plays a role in retrograde vesicular trafficking and Golgi-based quality control. The interaction and co-localization of kAE1 G701D with the γ-COPI subunit were demonstrated in human embryonic kidney (HEK-293T) cells by co-immunoprecipitation and immunofluorescence staining. Small interference RNA (siRNA) silencing of COPI expression in the transfected HEK-293T cells increased the cell surface expression of transgenic kAE1 G701D, as shown by immunofluorescence staining. Our data unveil the molecular mechanism of Golgi retention of kAE1 G701D and suggest that disruption of the COPI-kAE1 G701D interaction could be a therapeutic strategy to treat dRTA caused by this mutant. 2018-12-21T06:45:21Z 2019-03-14T08:02:50Z 2018-12-21T06:45:21Z 2019-03-14T08:02:50Z 2017-08-01 Article Biochemical Journal. Vol.474, No.15 (2017), 2573-2584 10.1042/BCJ20170088 14708728 02646021 2-s2.0-85026684776 https://repository.li.mahidol.ac.th/handle/123456789/41833 Mahidol University SCOPUS https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85026684776&origin=inward |
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Biochemistry, Genetics and Molecular Biology Natapol Duangtum Mutita Junking Suratchanee Phadngam Nunghathai Sawasdee Andrea Castiglioni Komgrid Charngkaew Thawornchai Limjindaporn Ciro Isidoro Pa Thai Yenchitsomanus γ-COPI mediates the retention of kAE1 G701D protein in Golgi apparatus – A mechanistic explanation of distal renal tubular acidosis associated with the G701D mutation |
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© 2017 The Author(s); published by Portland Press Limited on behalf of the Biochemical Society. Mutations of the solute carrier family 4 member 1 (SLC4A1) gene encoding kidney anion (chloride/bicarbonate ion) exchanger 1 (kAE1) can cause genetic distal renal tubular acidosis (dRTA). Different SLC4A1 mutations give rise to mutant kAE1 proteins with distinct defects in protein trafficking. The mutant kAE1 protein may be retained in endoplasmic reticulum (ER) or Golgi apparatus, or mis-targeted to the apical membrane, failing to display its function at the baso-lateral membrane. The ER-retained mutant kAE1 interacts with calnexin chaperone protein; disruption of this interaction permits the mutant kAE1 to reach the cell surface and display anion exchange activity. However, the mechanism of Golgi retention of mutant kAE1 G701D protein, which is otherwise functional, is still unclear. In the present study, we show that Golgi retention of kAE1 G701D is due to a stable interaction with the Golgi-resident protein, coat protein complex I (COPI), that plays a role in retrograde vesicular trafficking and Golgi-based quality control. The interaction and co-localization of kAE1 G701D with the γ-COPI subunit were demonstrated in human embryonic kidney (HEK-293T) cells by co-immunoprecipitation and immunofluorescence staining. Small interference RNA (siRNA) silencing of COPI expression in the transfected HEK-293T cells increased the cell surface expression of transgenic kAE1 G701D, as shown by immunofluorescence staining. Our data unveil the molecular mechanism of Golgi retention of kAE1 G701D and suggest that disruption of the COPI-kAE1 G701D interaction could be a therapeutic strategy to treat dRTA caused by this mutant. |
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Mahidol University |
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Mahidol University Natapol Duangtum Mutita Junking Suratchanee Phadngam Nunghathai Sawasdee Andrea Castiglioni Komgrid Charngkaew Thawornchai Limjindaporn Ciro Isidoro Pa Thai Yenchitsomanus |
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Natapol Duangtum Mutita Junking Suratchanee Phadngam Nunghathai Sawasdee Andrea Castiglioni Komgrid Charngkaew Thawornchai Limjindaporn Ciro Isidoro Pa Thai Yenchitsomanus |
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Natapol Duangtum |
title |
γ-COPI mediates the retention of kAE1 G701D protein in Golgi apparatus – A mechanistic explanation of distal renal tubular acidosis associated with the G701D mutation |
title_short |
γ-COPI mediates the retention of kAE1 G701D protein in Golgi apparatus – A mechanistic explanation of distal renal tubular acidosis associated with the G701D mutation |
title_full |
γ-COPI mediates the retention of kAE1 G701D protein in Golgi apparatus – A mechanistic explanation of distal renal tubular acidosis associated with the G701D mutation |
title_fullStr |
γ-COPI mediates the retention of kAE1 G701D protein in Golgi apparatus – A mechanistic explanation of distal renal tubular acidosis associated with the G701D mutation |
title_full_unstemmed |
γ-COPI mediates the retention of kAE1 G701D protein in Golgi apparatus – A mechanistic explanation of distal renal tubular acidosis associated with the G701D mutation |
title_sort |
γ-copi mediates the retention of kae1 g701d protein in golgi apparatus – a mechanistic explanation of distal renal tubular acidosis associated with the g701d mutation |
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2018 |
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https://repository.li.mahidol.ac.th/handle/123456789/41833 |
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1763489303431741440 |