Transient receptor potential canonical type 6 (TRPC6) O-GlcNAcylation at Threonine-221 plays potent role in channel regulation
Summary: Transient receptor potential canonical type 6 (TRPC6) is a non-voltage-gated channel that principally conducts calcium. Elevated channel activation contributes to fibrosis, hypertrophy, and proteinuria, often coupled to stimulation of nuclear factor of activated T-cells (NFAT). TRPC6 is pos...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-03-01
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| Series: | iScience |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2589004223003711 |
| _version_ | 1811154699854807040 |
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| author | Sumita Mishra Junfeng Ma Desirae McKoy Masayuki Sasaki Federica Farinelli Richard C. Page Mark J. Ranek Natasha Zachara David A. Kass |
| author_facet | Sumita Mishra Junfeng Ma Desirae McKoy Masayuki Sasaki Federica Farinelli Richard C. Page Mark J. Ranek Natasha Zachara David A. Kass |
| author_sort | Sumita Mishra |
| collection | DOAJ |
| description | Summary: Transient receptor potential canonical type 6 (TRPC6) is a non-voltage-gated channel that principally conducts calcium. Elevated channel activation contributes to fibrosis, hypertrophy, and proteinuria, often coupled to stimulation of nuclear factor of activated T-cells (NFAT). TRPC6 is post-translationally regulated, but a role for O-linked β-N-acetyl glucosamine (O-GlcNAcylation) as elevated by diabetes, is unknown. Here we show TRPC6 is constitutively O-GlcNAcylated at Ser14, Thr70, and Thr221 in the N-terminus ankryn-4 (AR4) and linker (LH1) domains. Mutagenesis to alanine reveals T221 as a critical controller of resting TRPC6 conductance, and associated NFAT activity and pro-hypertrophic signaling. T→A mutations at sites homologous in closely related TRPC3 and TRPC7 also increases their activity. Molecular modeling predicts interactions between Thr221-O-GlcNAc and Ser199, Glu200, and Glu246, and combined alanine substitutions of the latter similarly elevates resting NFAT activity. Thus, O-GlcNAcylated T221 and interactions with coordinating residues is required for normal TRPC6 channel conductance and NFAT activation. |
| first_indexed | 2024-04-10T04:20:28Z |
| format | Article |
| id | doaj.art-384a06f3f1a64bf0a65db1f0836c7956 |
| institution | Directory Open Access Journal |
| issn | 2589-0042 |
| language | English |
| last_indexed | 2024-04-10T04:20:28Z |
| publishDate | 2023-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | iScience |
| spelling | doaj.art-384a06f3f1a64bf0a65db1f0836c79562023-03-11T04:20:19ZengElsevieriScience2589-00422023-03-01263106294Transient receptor potential canonical type 6 (TRPC6) O-GlcNAcylation at Threonine-221 plays potent role in channel regulationSumita Mishra0Junfeng Ma1Desirae McKoy2Masayuki Sasaki3Federica Farinelli4Richard C. Page5Mark J. Ranek6Natasha Zachara7David A. Kass8Division of Cardiology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USADepartment of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University Medical Center, Washington, DC, USADivision of Cardiology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USADivision of Cardiology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USADivision of Cardiology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USADepartment of Chemistry and Biochemistry, Miami University, Oxford, OH, USADivision of Cardiology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USADepartment of Biological Chemistry, Department of Oncology, Johns Hopkins University, Baltimore, MD, USADivision of Cardiology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Department of Pharmacology and Molecular Sciences, Johns Hopkins University, Baltimore, MD, USA; Corresponding authorSummary: Transient receptor potential canonical type 6 (TRPC6) is a non-voltage-gated channel that principally conducts calcium. Elevated channel activation contributes to fibrosis, hypertrophy, and proteinuria, often coupled to stimulation of nuclear factor of activated T-cells (NFAT). TRPC6 is post-translationally regulated, but a role for O-linked β-N-acetyl glucosamine (O-GlcNAcylation) as elevated by diabetes, is unknown. Here we show TRPC6 is constitutively O-GlcNAcylated at Ser14, Thr70, and Thr221 in the N-terminus ankryn-4 (AR4) and linker (LH1) domains. Mutagenesis to alanine reveals T221 as a critical controller of resting TRPC6 conductance, and associated NFAT activity and pro-hypertrophic signaling. T→A mutations at sites homologous in closely related TRPC3 and TRPC7 also increases their activity. Molecular modeling predicts interactions between Thr221-O-GlcNAc and Ser199, Glu200, and Glu246, and combined alanine substitutions of the latter similarly elevates resting NFAT activity. Thus, O-GlcNAcylated T221 and interactions with coordinating residues is required for normal TRPC6 channel conductance and NFAT activation.http://www.sciencedirect.com/science/article/pii/S2589004223003711BiochemistryCellular physiologyCell biology |
| spellingShingle | Sumita Mishra Junfeng Ma Desirae McKoy Masayuki Sasaki Federica Farinelli Richard C. Page Mark J. Ranek Natasha Zachara David A. Kass Transient receptor potential canonical type 6 (TRPC6) O-GlcNAcylation at Threonine-221 plays potent role in channel regulation iScience Biochemistry Cellular physiology Cell biology |
| title | Transient receptor potential canonical type 6 (TRPC6) O-GlcNAcylation at Threonine-221 plays potent role in channel regulation |
| title_full | Transient receptor potential canonical type 6 (TRPC6) O-GlcNAcylation at Threonine-221 plays potent role in channel regulation |
| title_fullStr | Transient receptor potential canonical type 6 (TRPC6) O-GlcNAcylation at Threonine-221 plays potent role in channel regulation |
| title_full_unstemmed | Transient receptor potential canonical type 6 (TRPC6) O-GlcNAcylation at Threonine-221 plays potent role in channel regulation |
| title_short | Transient receptor potential canonical type 6 (TRPC6) O-GlcNAcylation at Threonine-221 plays potent role in channel regulation |
| title_sort | transient receptor potential canonical type 6 trpc6 o glcnacylation at threonine 221 plays potent role in channel regulation |
| topic | Biochemistry Cellular physiology Cell biology |
| url | http://www.sciencedirect.com/science/article/pii/S2589004223003711 |
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