Differential Internalization of Mammalian and Non-mammalian Gonadotropin-releasing Hormone Receptors: UNCOUPLING OF DYNAMIN-DEPENDENT INTERNALIZATION FROM MITOGEN-ACTIVATED PROTEIN KINASE SIGNALING

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dc.contributor.authorHislop, James N
dc.contributor.authorEverest, Helen M
dc.contributor.authorFlynn, Andrea
dc.contributor.authorHarding, Tom
dc.contributor.authorUney, James B
dc.contributor.authorTroskie, Brigitte E
dc.contributor.authorMillar, Robert P
dc.contributor.authorMcArdle, Craig A
dc.date.accessioned2021-10-08T07:20:47Z
dc.date.available2021-10-08T07:20:47Z
dc.date.issued2001
dc.description.abstractDesensitization and internalization of G-protein-coupled receptors can reflect receptor phosphorylation-dependent binding of beta-arrestin, which prevents G-protein activation and targets receptors for internalization via clathrin-coated vesicles. These can be pinched off by a dynamin collar, and proteins controlling receptor internalization can also mediate mitogen-activated protein kinase signaling. Gonadotropin-releasing hormone (GnRH) stimulates internalization of its receptors via clathrin-coated vesicles. Mammalian GnRH receptors (GnRH-Rs) are unique in that they lack C-terminal tails and do not rapidly desensitize, whereas non-mammalian GnRH-R have C-terminal tails and, where investigated, do rapidly desensitize and internalize. Using recombinant adenovirus expressing human and Xenopus GnRH-Rs we have explored the relationship between receptor internalization and mitogen-activated protein kinase signaling in HeLa cells with regulated tetracycline-controlled expression of wild-type or a dominant negative mutant (K44A) of dynamin. These receptors were phospholipase C-coupled and had appropriate ligand affinity and specificity. K44A dynamin expression did not alter human GnRH-R internalization but dramatically reduced internalization of Xenopus GnRH-R (and epidermal growth factor (EGF) receptor). Blockade of clathrin-mediated internalization (sucrose) abolished internalization of all three receptors. Both GnRH-Rs also mediated phosphorylation of ERK 2 and for both receptors, this was inhibited by K44A dynamin. The same was true for EGF- and protein kinase C-mediated ERK 2 phosphorylation. ERK 2 phosphorylation was also inhibited by a protein kinase C inhibitor but not affected by an EGF receptor tyrosine kinase inhibitor. We conclude that a) desensitizing and non-desensitizing GnRH-Rs are targeted for clathrin-coated vesicle-mediated internalization by functionally distinct mechanisms, b) GnRH-R signaling to ERK 2 is dynamin-dependent and c) this does not reflect a dependence on dynamin-dependent GnRH-R internalization.
dc.identifier.apacitationHislop, J. N., Everest, H. M., Flynn, A., Harding, T., Uney, J. B., Troskie, B. E., ... McArdle, C. A. (2001). Differential Internalization of Mammalian and Non-mammalian Gonadotropin-releasing Hormone Receptors: UNCOUPLING OF DYNAMIN-DEPENDENT INTERNALIZATION FROM MITOGEN-ACTIVATED PROTEIN KINASE SIGNALING. <i>The Journal of Biological Chemistry</i>, 276(43), 39685 - 39694. http://hdl.handle.net/11427/35003en_ZA
dc.identifier.chicagocitationHislop, James N, Helen M Everest, Andrea Flynn, Tom Harding, James B Uney, Brigitte E Troskie, Robert P Millar, and Craig A McArdle "Differential Internalization of Mammalian and Non-mammalian Gonadotropin-releasing Hormone Receptors: UNCOUPLING OF DYNAMIN-DEPENDENT INTERNALIZATION FROM MITOGEN-ACTIVATED PROTEIN KINASE SIGNALING." <i>The Journal of Biological Chemistry</i> 276, 43. (2001): 39685 - 39694. http://hdl.handle.net/11427/35003en_ZA
dc.identifier.citationHislop, J.N., Everest, H.M., Flynn, A., Harding, T., Uney, J.B., Troskie, B.E., Millar, R.P. & McArdle, C.A. et al. 2001. Differential Internalization of Mammalian and Non-mammalian Gonadotropin-releasing Hormone Receptors: UNCOUPLING OF DYNAMIN-DEPENDENT INTERNALIZATION FROM MITOGEN-ACTIVATED PROTEIN KINASE SIGNALING. <i>The Journal of Biological Chemistry.</i> 276(43):39685 - 39694. http://hdl.handle.net/11427/35003en_ZA
dc.identifier.issn0021-9258
dc.identifier.issn1083-351X
dc.identifier.ris TY - Journal Article AU - Hislop, James N AU - Everest, Helen M AU - Flynn, Andrea AU - Harding, Tom AU - Uney, James B AU - Troskie, Brigitte E AU - Millar, Robert P AU - McArdle, Craig A AB - Desensitization and internalization of G-protein-coupled receptors can reflect receptor phosphorylation-dependent binding of beta-arrestin, which prevents G-protein activation and targets receptors for internalization via clathrin-coated vesicles. These can be pinched off by a dynamin collar, and proteins controlling receptor internalization can also mediate mitogen-activated protein kinase signaling. Gonadotropin-releasing hormone (GnRH) stimulates internalization of its receptors via clathrin-coated vesicles. Mammalian GnRH receptors (GnRH-Rs) are unique in that they lack C-terminal tails and do not rapidly desensitize, whereas non-mammalian GnRH-R have C-terminal tails and, where investigated, do rapidly desensitize and internalize. Using recombinant adenovirus expressing human and Xenopus GnRH-Rs we have explored the relationship between receptor internalization and mitogen-activated protein kinase signaling in HeLa cells with regulated tetracycline-controlled expression of wild-type or a dominant negative mutant (K44A) of dynamin. These receptors were phospholipase C-coupled and had appropriate ligand affinity and specificity. K44A dynamin expression did not alter human GnRH-R internalization but dramatically reduced internalization of Xenopus GnRH-R (and epidermal growth factor (EGF) receptor). Blockade of clathrin-mediated internalization (sucrose) abolished internalization of all three receptors. Both GnRH-Rs also mediated phosphorylation of ERK 2 and for both receptors, this was inhibited by K44A dynamin. The same was true for EGF- and protein kinase C-mediated ERK 2 phosphorylation. ERK 2 phosphorylation was also inhibited by a protein kinase C inhibitor but not affected by an EGF receptor tyrosine kinase inhibitor. We conclude that a) desensitizing and non-desensitizing GnRH-Rs are targeted for clathrin-coated vesicle-mediated internalization by functionally distinct mechanisms, b) GnRH-R signaling to ERK 2 is dynamin-dependent and c) this does not reflect a dependence on dynamin-dependent GnRH-R internalization. DA - 2001 DB - OpenUCT DP - University of Cape Town IS - 43 J1 - The Journal of Biological Chemistry LK - https://open.uct.ac.za PY - 2001 SM - 0021-9258 SM - 1083-351X T1 - Differential Internalization of Mammalian and Non-mammalian Gonadotropin-releasing Hormone Receptors: UNCOUPLING OF DYNAMIN-DEPENDENT INTERNALIZATION FROM MITOGEN-ACTIVATED PROTEIN KINASE SIGNALING TI - Differential Internalization of Mammalian and Non-mammalian Gonadotropin-releasing Hormone Receptors: UNCOUPLING OF DYNAMIN-DEPENDENT INTERNALIZATION FROM MITOGEN-ACTIVATED PROTEIN KINASE SIGNALING UR - http://hdl.handle.net/11427/35003 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/35003
dc.identifier.vancouvercitationHislop JN, Everest HM, Flynn A, Harding T, Uney JB, Troskie BE, et al. Differential Internalization of Mammalian and Non-mammalian Gonadotropin-releasing Hormone Receptors: UNCOUPLING OF DYNAMIN-DEPENDENT INTERNALIZATION FROM MITOGEN-ACTIVATED PROTEIN KINASE SIGNALING. The Journal of Biological Chemistry. 2001;276(43):39685 - 39694. http://hdl.handle.net/11427/35003.en_ZA
dc.language.isoeng
dc.publisher.departmentDivision of Medical Biochemistry
dc.publisher.facultyFaculty of Health Sciences
dc.sourceThe Journal of Biological Chemistry
dc.source.journalissue43
dc.source.journalvolume276
dc.source.pagination39685 - 39694
dc.source.urihttps://dx.doi.org/10.1074/jbc.M104542200
dc.subject.otherAnimals
dc.subject.otherDynamins
dc.subject.otherEndocytosis
dc.subject.otherGTP Phosphohydrolases
dc.subject.otherHumans
dc.subject.otherInositol Phosphates
dc.subject.otherMitogen-Activated Protein Kinase 1
dc.subject.otherMitogen-Activated Protein Kinases
dc.subject.otherReceptors, LHRH
dc.subject.otherRecombinant Proteins
dc.subject.otherSignal Transduction
dc.subject.otherSpecies Specificity
dc.subject.otherInositol Phosphates
dc.subject.otherReceptors, LHRH
dc.subject.otherRecombinant Proteins
dc.titleDifferential Internalization of Mammalian and Non-mammalian Gonadotropin-releasing Hormone Receptors: UNCOUPLING OF DYNAMIN-DEPENDENT INTERNALIZATION FROM MITOGEN-ACTIVATED PROTEIN KINASE SIGNALING
dc.typeJournal Article
uct.type.publicationResearch
uct.type.resourceJournal Article
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