Internalization of gonadotropin-releasing hormone receptors (GnRHRs): does arrestin binding to the C-terminal tail target GnRHRs for dynamin-dependent internalization?

James N Hislop, Christopher J Caunt, Kathleen R Sedgley, Eammon Kelly, Stuart Mundell, Lisa D Green, Craig A McArdle

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Activation of seven-transmembrane receptors is typically followed by desensitization and arrestin-dependent internalization via vesicles that are pinched off by a dynamin collar. Arrestins also scaffold Src, which mediates dynamin-dependent internalization of beta2-adrenergic receptors. Type I mammalian gonadotropin-releasing hormone receptors (GnRHRs) do not rapidly desensitize or internalize (characteristics attributed to their unique lack of C-terminal tails) whereas non-mammalian GnRHRs (that have C-terminal tails) are rapidly internalized and desensitized. Moreover, internalization of Xenopus (X) GnRHRs is dynamin-dependent whereas that of human (h) GnRHRs is not, raising the possibility that binding of arrestin to the C-terminal tails of GnRHRs targets them to the dynamin-dependent internalization pathway. To test this we have compared wild-type GnRHRs with chimeric receptors (XGnRHR C-terminal tail added to the hGnRHR alone (h.XtGnRHR) or with exchange of the third intracellular loops (h.Xl.XtGnRHR)). We show that adding the XGnRHR C-terminal tail facilitates arrestin- and dynamin-dependent internalization as well as arrestin/green fluorescent protein translocation, but Src (or mitogen-activated protein kinase/extracellular-signal-regulated kinase kinase) inhibition does not slow internalization, and h.XtGnRHR internalization is slower than that of the hGnRHR. Moreover, arrestin expression increased XGnRHR internalization even when dynamin was inhibited and h.Xl.XtGnRHR underwent rapid arrestin-dependent internalization without signaling to G(q/11). Thus, although the C-terminal tail can direct GnRHRs for arrestin- and dynamin-dependent internalization, this effect is not dependent on Src activation and arrestin can also facilitate dynamin-independent internalization.
Original languageEnglish
Pages (from-to)177-89
Number of pages13
JournalJournal of Molecular Endocrinology
Volume35
Issue number1
DOIs
Publication statusPublished - Aug 2005

Fingerprint

Dynamins
LHRH Receptors
Arrestin
Tail
Arrestins
Extracellular Signal-Regulated MAP Kinases
Protein Transport
Xenopus
Green Fluorescent Proteins
Mitogen-Activated Protein Kinases
Adrenergic Receptors
Phosphotransferases

Keywords

  • Animals
  • Arrestin
  • Base Sequence
  • Binding Sites
  • Binding, Competitive
  • DNA, Complementary
  • Dynamins
  • Green Fluorescent Proteins
  • HeLa Cells
  • Humans
  • Kinetics
  • Protein Binding
  • Protein Engineering
  • Receptors, LHRH
  • Recombinant Fusion Proteins
  • Xenopus

Cite this

Internalization of gonadotropin-releasing hormone receptors (GnRHRs) : does arrestin binding to the C-terminal tail target GnRHRs for dynamin-dependent internalization? / Hislop, James N; Caunt, Christopher J; Sedgley, Kathleen R; Kelly, Eammon; Mundell, Stuart; Green, Lisa D; McArdle, Craig A.

In: Journal of Molecular Endocrinology, Vol. 35, No. 1, 08.2005, p. 177-89.

Research output: Contribution to journalArticle

Hislop, James N ; Caunt, Christopher J ; Sedgley, Kathleen R ; Kelly, Eammon ; Mundell, Stuart ; Green, Lisa D ; McArdle, Craig A. / Internalization of gonadotropin-releasing hormone receptors (GnRHRs) : does arrestin binding to the C-terminal tail target GnRHRs for dynamin-dependent internalization?. In: Journal of Molecular Endocrinology. 2005 ; Vol. 35, No. 1. pp. 177-89.
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AU - Hislop, James N

AU - Caunt, Christopher J

AU - Sedgley, Kathleen R

AU - Kelly, Eammon

AU - Mundell, Stuart

AU - Green, Lisa D

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N2 - Activation of seven-transmembrane receptors is typically followed by desensitization and arrestin-dependent internalization via vesicles that are pinched off by a dynamin collar. Arrestins also scaffold Src, which mediates dynamin-dependent internalization of beta2-adrenergic receptors. Type I mammalian gonadotropin-releasing hormone receptors (GnRHRs) do not rapidly desensitize or internalize (characteristics attributed to their unique lack of C-terminal tails) whereas non-mammalian GnRHRs (that have C-terminal tails) are rapidly internalized and desensitized. Moreover, internalization of Xenopus (X) GnRHRs is dynamin-dependent whereas that of human (h) GnRHRs is not, raising the possibility that binding of arrestin to the C-terminal tails of GnRHRs targets them to the dynamin-dependent internalization pathway. To test this we have compared wild-type GnRHRs with chimeric receptors (XGnRHR C-terminal tail added to the hGnRHR alone (h.XtGnRHR) or with exchange of the third intracellular loops (h.Xl.XtGnRHR)). We show that adding the XGnRHR C-terminal tail facilitates arrestin- and dynamin-dependent internalization as well as arrestin/green fluorescent protein translocation, but Src (or mitogen-activated protein kinase/extracellular-signal-regulated kinase kinase) inhibition does not slow internalization, and h.XtGnRHR internalization is slower than that of the hGnRHR. Moreover, arrestin expression increased XGnRHR internalization even when dynamin was inhibited and h.Xl.XtGnRHR underwent rapid arrestin-dependent internalization without signaling to G(q/11). Thus, although the C-terminal tail can direct GnRHRs for arrestin- and dynamin-dependent internalization, this effect is not dependent on Src activation and arrestin can also facilitate dynamin-independent internalization.

AB - Activation of seven-transmembrane receptors is typically followed by desensitization and arrestin-dependent internalization via vesicles that are pinched off by a dynamin collar. Arrestins also scaffold Src, which mediates dynamin-dependent internalization of beta2-adrenergic receptors. Type I mammalian gonadotropin-releasing hormone receptors (GnRHRs) do not rapidly desensitize or internalize (characteristics attributed to their unique lack of C-terminal tails) whereas non-mammalian GnRHRs (that have C-terminal tails) are rapidly internalized and desensitized. Moreover, internalization of Xenopus (X) GnRHRs is dynamin-dependent whereas that of human (h) GnRHRs is not, raising the possibility that binding of arrestin to the C-terminal tails of GnRHRs targets them to the dynamin-dependent internalization pathway. To test this we have compared wild-type GnRHRs with chimeric receptors (XGnRHR C-terminal tail added to the hGnRHR alone (h.XtGnRHR) or with exchange of the third intracellular loops (h.Xl.XtGnRHR)). We show that adding the XGnRHR C-terminal tail facilitates arrestin- and dynamin-dependent internalization as well as arrestin/green fluorescent protein translocation, but Src (or mitogen-activated protein kinase/extracellular-signal-regulated kinase kinase) inhibition does not slow internalization, and h.XtGnRHR internalization is slower than that of the hGnRHR. Moreover, arrestin expression increased XGnRHR internalization even when dynamin was inhibited and h.Xl.XtGnRHR underwent rapid arrestin-dependent internalization without signaling to G(q/11). Thus, although the C-terminal tail can direct GnRHRs for arrestin- and dynamin-dependent internalization, this effect is not dependent on Src activation and arrestin can also facilitate dynamin-independent internalization.

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KW - Base Sequence

KW - Binding Sites

KW - Binding, Competitive

KW - DNA, Complementary

KW - Dynamins

KW - Green Fluorescent Proteins

KW - HeLa Cells

KW - Humans

KW - Kinetics

KW - Protein Binding

KW - Protein Engineering

KW - Receptors, LHRH

KW - Recombinant Fusion Proteins

KW - Xenopus

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JO - Journal of Molecular Endocrinology

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