Phosphorylation of the dimeric cytoplasmic domain of the phytosulfokine receptor, PSKR1
Phytosulfokines (PSKs) are plant peptide hormones that co-regulate plant growth, differentiation and defense responses. PSKs signal through a plasma membrane localized leucine-rich repeat receptor-like kinase (phytosulfokine receptor 1, PSKR1) that also contains a functional cytosolic guanylate cycl...
Saved in:
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Portland Press
2021
|
| Subjects: | |
| Online Access: | https://portlandpress.com/biochemj/article-abstract/473/19/3081/49606/Phosphorylation-of-the-dimeric-cytoplasmic-domain?redirectedFrom=fulltext https://doi.org/10.1042/BCJ20160593 http://hdl.handle.net/11408/4438 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1779905668837277696 |
|---|---|
| author | Muleya, Victor Marondedze, Claudius Wheeler, Janet I. Thomas, Ludivine Mok, Yee-Fong Griffin, Michael D.W. Manallack, David T. Kwezi, Lusisizwe Lilley, Kathryn S. Gehring, Christoph Irving, Helen R. |
| author_facet | Muleya, Victor Marondedze, Claudius Wheeler, Janet I. Thomas, Ludivine Mok, Yee-Fong Griffin, Michael D.W. Manallack, David T. Kwezi, Lusisizwe Lilley, Kathryn S. Gehring, Christoph Irving, Helen R. |
| author_sort | Muleya, Victor |
| collection | DSpace |
| description | Phytosulfokines (PSKs) are plant peptide hormones that co-regulate plant growth, differentiation and defense responses. PSKs signal through a plasma membrane localized leucine-rich repeat receptor-like kinase (phytosulfokine receptor 1, PSKR1) that also contains a functional cytosolic guanylate cyclase with its cyclase catalytic center embedded within the kinase domain. To functionally characterize this novel type of overlapping dual catalytic function, we investigated the phosphorylation of PSKR1 in vitro. Tandem mass spectrometry of the cytoplasmic domain of PSKR1 (PSKR1cd) revealed at least 11 phosphorylation sites (8 serines, 2 threonines and 1 tyrosine) within the PSKR1cd. Phosphomimetic mutations of three serine residues (Ser686, Ser696 and Ser698) in tandem at the juxta-membrane position resulted in enhanced kinase activity in the on-mutant that was suppressed in the off-mutant, but both mutations reduced guanylate cyclase activity. Both the on and off phosphomimetic mutations of the phosphotyrosine (Tyr888) residue in the activation loop suppressed kinase activity, while neither mutation affected guanylate cyclase activity. Size exclusion and analytical ultracentrifugation analysis of the PSKR1cd suggest that it is reversibly dimeric in solution, which was further confirmed by biflourescence complementation. Taken together, these data suggest that in this novel type of receptor domain architecture, specific phosphorylation and dimerization are possibly essential mechanisms for ligand-mediated catalysis and signaling. |
| format | Article |
| id | ir-11408-4438 |
| institution | My University |
| language | English |
| publishDate | 2021 |
| publisher | Portland Press |
| record_format | dspace |
| spelling | ir-11408-44382022-10-15T20:45:40Z Phosphorylation of the dimeric cytoplasmic domain of the phytosulfokine receptor, PSKR1 Muleya, Victor Marondedze, Claudius Wheeler, Janet I. Thomas, Ludivine Mok, Yee-Fong Griffin, Michael D.W. Manallack, David T. Kwezi, Lusisizwe Lilley, Kathryn S. Gehring, Christoph Irving, Helen R. Dimerization Dual-specificity kinase Computational Biology Phytosulfokines (PSKs) are plant peptide hormones that co-regulate plant growth, differentiation and defense responses. PSKs signal through a plasma membrane localized leucine-rich repeat receptor-like kinase (phytosulfokine receptor 1, PSKR1) that also contains a functional cytosolic guanylate cyclase with its cyclase catalytic center embedded within the kinase domain. To functionally characterize this novel type of overlapping dual catalytic function, we investigated the phosphorylation of PSKR1 in vitro. Tandem mass spectrometry of the cytoplasmic domain of PSKR1 (PSKR1cd) revealed at least 11 phosphorylation sites (8 serines, 2 threonines and 1 tyrosine) within the PSKR1cd. Phosphomimetic mutations of three serine residues (Ser686, Ser696 and Ser698) in tandem at the juxta-membrane position resulted in enhanced kinase activity in the on-mutant that was suppressed in the off-mutant, but both mutations reduced guanylate cyclase activity. Both the on and off phosphomimetic mutations of the phosphotyrosine (Tyr888) residue in the activation loop suppressed kinase activity, while neither mutation affected guanylate cyclase activity. Size exclusion and analytical ultracentrifugation analysis of the PSKR1cd suggest that it is reversibly dimeric in solution, which was further confirmed by biflourescence complementation. Taken together, these data suggest that in this novel type of receptor domain architecture, specific phosphorylation and dimerization are possibly essential mechanisms for ligand-mediated catalysis and signaling. 2021-06-09T13:34:50Z 2021-06-09T13:34:50Z 2016 Article 0264-6021 1470-8728 https://portlandpress.com/biochemj/article-abstract/473/19/3081/49606/Phosphorylation-of-the-dimeric-cytoplasmic-domain?redirectedFrom=fulltext https://doi.org/10.1042/BCJ20160593 http://hdl.handle.net/11408/4438 en Biochemical Journal;Vol. 473; No. 19: p. 3081-3098 open Portland Press |
| spellingShingle | Dimerization Dual-specificity kinase Computational Biology Muleya, Victor Marondedze, Claudius Wheeler, Janet I. Thomas, Ludivine Mok, Yee-Fong Griffin, Michael D.W. Manallack, David T. Kwezi, Lusisizwe Lilley, Kathryn S. Gehring, Christoph Irving, Helen R. Phosphorylation of the dimeric cytoplasmic domain of the phytosulfokine receptor, PSKR1 |
| title | Phosphorylation of the dimeric cytoplasmic domain of the phytosulfokine receptor, PSKR1 |
| title_full | Phosphorylation of the dimeric cytoplasmic domain of the phytosulfokine receptor, PSKR1 |
| title_fullStr | Phosphorylation of the dimeric cytoplasmic domain of the phytosulfokine receptor, PSKR1 |
| title_full_unstemmed | Phosphorylation of the dimeric cytoplasmic domain of the phytosulfokine receptor, PSKR1 |
| title_short | Phosphorylation of the dimeric cytoplasmic domain of the phytosulfokine receptor, PSKR1 |
| title_sort | phosphorylation of the dimeric cytoplasmic domain of the phytosulfokine receptor, pskr1 |
| topic | Dimerization Dual-specificity kinase Computational Biology |
| url | https://portlandpress.com/biochemj/article-abstract/473/19/3081/49606/Phosphorylation-of-the-dimeric-cytoplasmic-domain?redirectedFrom=fulltext https://doi.org/10.1042/BCJ20160593 http://hdl.handle.net/11408/4438 |
| work_keys_str_mv | AT muleyavictor phosphorylationofthedimericcytoplasmicdomainofthephytosulfokinereceptorpskr1 AT marondedzeclaudius phosphorylationofthedimericcytoplasmicdomainofthephytosulfokinereceptorpskr1 AT wheelerjaneti phosphorylationofthedimericcytoplasmicdomainofthephytosulfokinereceptorpskr1 AT thomasludivine phosphorylationofthedimericcytoplasmicdomainofthephytosulfokinereceptorpskr1 AT mokyeefong phosphorylationofthedimericcytoplasmicdomainofthephytosulfokinereceptorpskr1 AT griffinmichaeldw phosphorylationofthedimericcytoplasmicdomainofthephytosulfokinereceptorpskr1 AT manallackdavidt phosphorylationofthedimericcytoplasmicdomainofthephytosulfokinereceptorpskr1 AT kwezilusisizwe phosphorylationofthedimericcytoplasmicdomainofthephytosulfokinereceptorpskr1 AT lilleykathryns phosphorylationofthedimericcytoplasmicdomainofthephytosulfokinereceptorpskr1 AT gehringchristoph phosphorylationofthedimericcytoplasmicdomainofthephytosulfokinereceptorpskr1 AT irvinghelenr phosphorylationofthedimericcytoplasmicdomainofthephytosulfokinereceptorpskr1 |