Leu-574 of human HIF-1alpha is a molecular determinant of prolyl hydroxylation.
Hypoxia-inducible factor (HIF)-1alpha, a master regulator of oxygen homeostasis, regulates genes crucial for cell growth and survival. In normoxia, HIF-1alpha is constantly degraded via the ubiquitin-proteasome pathway. The von Hippel-Lindau (VHL) E3 ubiquitin ligase binds HIF-1alpha through specifi...
| Váldodahkkit: | , , , , , |
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| Materiálatiipa: | Journal article |
| Giella: | English |
| Almmustuhtton: |
2004
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| _version_ | 1826294095103918080 |
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| author | Kageyama, Y Koshiji, M To, K Tian, Y Ratcliffe, P Huang, L |
| author_facet | Kageyama, Y Koshiji, M To, K Tian, Y Ratcliffe, P Huang, L |
| author_sort | Kageyama, Y |
| collection | OXFORD |
| description | Hypoxia-inducible factor (HIF)-1alpha, a master regulator of oxygen homeostasis, regulates genes crucial for cell growth and survival. In normoxia, HIF-1alpha is constantly degraded via the ubiquitin-proteasome pathway. The von Hippel-Lindau (VHL) E3 ubiquitin ligase binds HIF-1alpha through specific recognition of hydroxylated Pro-402 or Pro-564, both of which are modified by the oxygen-dependent HIF prolyl hydroxylases (PHDs/HPHs). Despite the identification of a conserved Leu-X-X-Leu-Ala-Pro motif, the molecular requirement of HIF-1alpha for PHDs/HPHs binding remains elusive. Recently, we demonstrated that Leu-574 of human HIF-1alpha--10 residues downstream of Pro-564--is essential for VHL recognition. We show here that the role of Leu-574 is to recruit PHD2/HPH2 for Pro-564 hydroxylation. An antibody specific for hydroxylated Pro-564 has been used to determine the hydroxylation status; mutation or deletion of Leu-574 results in a significant decrease in the ratio of the hydroxylated HIF-1alpha to the total amount. The nine-residue spacing between Pro-564 and Leu-574 is not obligatory for prolyl hydroxylation. Furthermore, mutation of Leu-574 disrupts the binding of PHD2/HPH2, a key prolyl hydroxylase for oxygen-dependent proteolysis of HIF-1alpha. Hence, our findings indicate that Leu-574 is essential for recruiting PHD2/HPH2, thereby providing a molecular basis for modulating HIF-1alpha activity. |
| first_indexed | 2024-03-07T03:40:20Z |
| format | Journal article |
| id | oxford-uuid:bdaae5df-fd18-4e76-8314-2591de6d74cd |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T03:40:20Z |
| publishDate | 2004 |
| record_format | dspace |
| spelling | oxford-uuid:bdaae5df-fd18-4e76-8314-2591de6d74cd2022-03-27T05:33:38ZLeu-574 of human HIF-1alpha is a molecular determinant of prolyl hydroxylation.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:bdaae5df-fd18-4e76-8314-2591de6d74cdEnglishSymplectic Elements at Oxford2004Kageyama, YKoshiji, MTo, KTian, YRatcliffe, PHuang, LHypoxia-inducible factor (HIF)-1alpha, a master regulator of oxygen homeostasis, regulates genes crucial for cell growth and survival. In normoxia, HIF-1alpha is constantly degraded via the ubiquitin-proteasome pathway. The von Hippel-Lindau (VHL) E3 ubiquitin ligase binds HIF-1alpha through specific recognition of hydroxylated Pro-402 or Pro-564, both of which are modified by the oxygen-dependent HIF prolyl hydroxylases (PHDs/HPHs). Despite the identification of a conserved Leu-X-X-Leu-Ala-Pro motif, the molecular requirement of HIF-1alpha for PHDs/HPHs binding remains elusive. Recently, we demonstrated that Leu-574 of human HIF-1alpha--10 residues downstream of Pro-564--is essential for VHL recognition. We show here that the role of Leu-574 is to recruit PHD2/HPH2 for Pro-564 hydroxylation. An antibody specific for hydroxylated Pro-564 has been used to determine the hydroxylation status; mutation or deletion of Leu-574 results in a significant decrease in the ratio of the hydroxylated HIF-1alpha to the total amount. The nine-residue spacing between Pro-564 and Leu-574 is not obligatory for prolyl hydroxylation. Furthermore, mutation of Leu-574 disrupts the binding of PHD2/HPH2, a key prolyl hydroxylase for oxygen-dependent proteolysis of HIF-1alpha. Hence, our findings indicate that Leu-574 is essential for recruiting PHD2/HPH2, thereby providing a molecular basis for modulating HIF-1alpha activity. |
| spellingShingle | Kageyama, Y Koshiji, M To, K Tian, Y Ratcliffe, P Huang, L Leu-574 of human HIF-1alpha is a molecular determinant of prolyl hydroxylation. |
| title | Leu-574 of human HIF-1alpha is a molecular determinant of prolyl hydroxylation. |
| title_full | Leu-574 of human HIF-1alpha is a molecular determinant of prolyl hydroxylation. |
| title_fullStr | Leu-574 of human HIF-1alpha is a molecular determinant of prolyl hydroxylation. |
| title_full_unstemmed | Leu-574 of human HIF-1alpha is a molecular determinant of prolyl hydroxylation. |
| title_short | Leu-574 of human HIF-1alpha is a molecular determinant of prolyl hydroxylation. |
| title_sort | leu 574 of human hif 1alpha is a molecular determinant of prolyl hydroxylation |
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