Structure-function analysis of β-arrestin Kurtz reveals a critical role of receptor interactions in downregulation of GPCR signaling in vivo.
Amino Acid Motifs
Animals
Arrestins
/ chemistry
Down-Regulation
Drosophila
/ embryology
Drosophila Proteins
/ chemistry
Female
Gastrulation
Male
Models, Molecular
Protein Conformation
Receptors, G-Protein-Coupled
/ metabolism
Signal Transduction
Structure-Activity Relationship
Wings, Animal
/ embryology
Arrestin
Drosophila
Epithelial morphogenesis
G protein coupled receptor
Gastrulation
Kurtz
Journal
Developmental biology
ISSN: 1095-564X
Titre abrégé: Dev Biol
Pays: United States
ID NLM: 0372762
Informations de publication
Date de publication:
15 11 2019
15 11 2019
Historique:
received:
19
05
2019
revised:
15
06
2019
accepted:
16
07
2019
pubmed:
22
7
2019
medline:
15
5
2020
entrez:
21
7
2019
Statut:
ppublish
Résumé
Arrestins control signaling via the G protein coupled receptors (GPCRs), serving as both signal terminators and transducers. Previous studies identified several structural elements in arrestins that contribute to their functions as GPCR regulators. However, the importance of these elements in vivo is unclear, and the developmental roles of arrestins are not well understood. We carried out an in vivo structure-function analysis of Kurtz (Krz), the single ortholog of mammalian β-arrestins in the Drosophila genome. A combination of Krz mutations affecting the GPCR-phosphosensing and receptor core-binding ("finger loop") functions (Krz-KKVL/A) resulted in a complete loss of Krz activity during development. Endosome recruitment and bioluminescence resonance energy transfer (BRET) assays revealed that the KKVL/A mutations abolished the GPCR-binding ability of Krz. We found that the isolated "finger loop" mutation (Krz-VL/A), while having a negligible effect on GPCR internalization, severely affected Krz function, suggesting that tight receptor interactions are necessary for proper termination of signaling in vivo. Genetic analysis as well as live imaging demonstrated that mutations in Krz led to hyperactivity of the GPCR Mist (also known as Mthl1), which is activated by its ligand Folded gastrulation (Fog) and is responsible for cellular contractility and epithelial morphogenesis. Krz mutations affected two developmental events that are under the control of Fog-Mist signaling: gastrulation and morphogenesis of the wing. Overall, our data reveal the functional importance in vivo of direct β-arrestin/GPCR binding, which is mediated by the recognition of the phosphorylated receptor tail and receptor core interaction. These Krz-GPCR interactions are critical for setting the correct level of Fog-Mist signaling during epithelial morphogenesis.
Identifiants
pubmed: 31325455
pii: S0012-1606(19)30314-8
doi: 10.1016/j.ydbio.2019.07.013
pmc: PMC6842422
mid: NIHMS1535480
pii:
doi:
Substances chimiques
Arrestins
0
Drosophila Proteins
0
FOG protein, Drosophila
0
Receptors, G-Protein-Coupled
0
krz protein, Drosophila
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
409-419Subventions
Organisme : NIGMS NIH HHS
ID : R15 GM097727
Pays : United States
Informations de copyright
Copyright © 2019 Elsevier Inc. All rights reserved.
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