Endothelial PKA activity regulates angiogenesis by limiting autophagy through phosphorylation of ATG16L1
The cAMP-dependent protein kinase A (PKA) regulates various cellular functions in health and disease. In endothelial cells PKA activity promotes vessel maturation and limits tip cell formation. Here, we used a chemical genetic screen to identify endothelial-specific direct substrates of PKA in human...
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2019-10-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/46380 |
| _version_ | 1818023838064050176 |
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| author | Xiaocheng Zhao Pavel Nedvetsky Fabio Stanchi Anne-Clemence Vion Oliver Popp Kerstin Zühlke Gunnar Dittmar Enno Klussmann Holger Gerhardt |
| author_facet | Xiaocheng Zhao Pavel Nedvetsky Fabio Stanchi Anne-Clemence Vion Oliver Popp Kerstin Zühlke Gunnar Dittmar Enno Klussmann Holger Gerhardt |
| author_sort | Xiaocheng Zhao |
| collection | DOAJ |
| description | The cAMP-dependent protein kinase A (PKA) regulates various cellular functions in health and disease. In endothelial cells PKA activity promotes vessel maturation and limits tip cell formation. Here, we used a chemical genetic screen to identify endothelial-specific direct substrates of PKA in human umbilical vein endothelial cells (HUVEC) that may mediate these effects. Amongst several candidates, we identified ATG16L1, a regulator of autophagy, as novel target of PKA. Biochemical validation, mass spectrometry and peptide spot arrays revealed that PKA phosphorylates ATG16L1α at Ser268 and ATG16L1β at Ser269, driving phosphorylation-dependent degradation of ATG16L1 protein. Reducing PKA activity increased ATG16L1 protein levels and endothelial autophagy. Mouse in vivo genetics and pharmacological experiments demonstrated that autophagy inhibition partially rescues vascular hypersprouting caused by PKA deficiency. Together these results indicate that endothelial PKA activity mediates a critical switch from active sprouting to quiescence in part through phosphorylation of ATG16L1, which in turn reduces endothelial autophagy. |
| first_indexed | 2024-12-10T03:50:40Z |
| format | Article |
| id | doaj.art-d866cab8e84a4592975aa952d912b1a6 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-12-10T03:50:40Z |
| publishDate | 2019-10-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-d866cab8e84a4592975aa952d912b1a62022-12-22T02:03:16ZengeLife Sciences Publications LtdeLife2050-084X2019-10-01810.7554/eLife.46380Endothelial PKA activity regulates angiogenesis by limiting autophagy through phosphorylation of ATG16L1Xiaocheng Zhao0https://orcid.org/0000-0002-4048-6813Pavel Nedvetsky1Fabio Stanchi2Anne-Clemence Vion3https://orcid.org/0000-0002-2788-2512Oliver Popp4Kerstin Zühlke5Gunnar Dittmar6Enno Klussmann7https://orcid.org/0000-0003-4004-5003Holger Gerhardt8https://orcid.org/0000-0002-3030-0384Vascular Patterning Laboratory, Center for Cancer Biology, VIB, Leuven, Belgium; Vascular Patterning Laboratory, Center for Cancer Biology, Department of Oncology, VIB, Leuven, BelgiumVascular Patterning Laboratory, Center for Cancer Biology, VIB, Leuven, Belgium; Vascular Patterning Laboratory, Center for Cancer Biology, Department of Oncology, VIB, Leuven, Belgium; Medical Cell Biology, Medical Clinic D, University Hospital Münster, Münster, GermanyVascular Patterning Laboratory, Center for Cancer Biology, VIB, Leuven, Belgium; Vascular Patterning Laboratory, Center for Cancer Biology, Department of Oncology, VIB, Leuven, BelgiumIntegrative Vascular Biology Lab, Max-Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany; INSERM UMR-970, Paris Cardiovascular Research Center, Paris Descartes University, Paris, FranceProteomics, Max-Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, GermanyAnchored Signaling Lab, Max-Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, GermanyProteomics, Max-Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany; CRP Santé · Department of Oncology, LIH Luxembourg Institute of Health, Luxembourg, LuxembourgAnchored Signaling Lab, Max-Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany; DZHK (German Center for Cardiovascular Research), Berlin, GermanyVascular Patterning Laboratory, Center for Cancer Biology, VIB, Leuven, Belgium; Vascular Patterning Laboratory, Center for Cancer Biology, Department of Oncology, VIB, Leuven, Belgium; Integrative Vascular Biology Lab, Max-Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany; DZHK (German Center for Cardiovascular Research), Berlin, Germany; Berlin Institute of Health (BIH), Berlin, GermanyThe cAMP-dependent protein kinase A (PKA) regulates various cellular functions in health and disease. In endothelial cells PKA activity promotes vessel maturation and limits tip cell formation. Here, we used a chemical genetic screen to identify endothelial-specific direct substrates of PKA in human umbilical vein endothelial cells (HUVEC) that may mediate these effects. Amongst several candidates, we identified ATG16L1, a regulator of autophagy, as novel target of PKA. Biochemical validation, mass spectrometry and peptide spot arrays revealed that PKA phosphorylates ATG16L1α at Ser268 and ATG16L1β at Ser269, driving phosphorylation-dependent degradation of ATG16L1 protein. Reducing PKA activity increased ATG16L1 protein levels and endothelial autophagy. Mouse in vivo genetics and pharmacological experiments demonstrated that autophagy inhibition partially rescues vascular hypersprouting caused by PKA deficiency. Together these results indicate that endothelial PKA activity mediates a critical switch from active sprouting to quiescence in part through phosphorylation of ATG16L1, which in turn reduces endothelial autophagy.https://elifesciences.org/articles/46380angiogenesisautophagychemical geneticsprotein kinase A |
| spellingShingle | Xiaocheng Zhao Pavel Nedvetsky Fabio Stanchi Anne-Clemence Vion Oliver Popp Kerstin Zühlke Gunnar Dittmar Enno Klussmann Holger Gerhardt Endothelial PKA activity regulates angiogenesis by limiting autophagy through phosphorylation of ATG16L1 eLife angiogenesis autophagy chemical genetics protein kinase A |
| title | Endothelial PKA activity regulates angiogenesis by limiting autophagy through phosphorylation of ATG16L1 |
| title_full | Endothelial PKA activity regulates angiogenesis by limiting autophagy through phosphorylation of ATG16L1 |
| title_fullStr | Endothelial PKA activity regulates angiogenesis by limiting autophagy through phosphorylation of ATG16L1 |
| title_full_unstemmed | Endothelial PKA activity regulates angiogenesis by limiting autophagy through phosphorylation of ATG16L1 |
| title_short | Endothelial PKA activity regulates angiogenesis by limiting autophagy through phosphorylation of ATG16L1 |
| title_sort | endothelial pka activity regulates angiogenesis by limiting autophagy through phosphorylation of atg16l1 |
| topic | angiogenesis autophagy chemical genetics protein kinase A |
| url | https://elifesciences.org/articles/46380 |
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