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Regulation of transport of the dopamine D1 receptor by a new membrane-associated ER protein

Nature Cell Biology volume 3, pages 492–498 (2001)Cite this article

Abstract

Many structural determinants for G protein-coupled receptor (GPCR) functions have been defined, but little is known concerning the regulation of their transport from the endoplasmic reticulum (ER) to the cell surface. Here we show that a carboxy-terminal hydrophobic motif, FxxxFxxxF, which is highly conserved among GPCRs, functions independently as an ER-export signal for the dopamine D1 receptor. A newly identified ER-membrane-associated protein, DRiP78, binds to this motif. Overexpression or sequestration of DRiP78 leads to retention of D1 receptors in the ER, reduced ligand binding, and a slowdown in the kinetics of receptor glycosylation. Our results indicate that DRiP78 may regulate the transport of a GPCR by binding to a specific ER-export signal.

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Figure 1: Transport of wild-type and mutant D1 receptors.
Figure 2: Localization and pulse–chase analysis of CD8–D1(wt)ct and CD8–D1(FA)ct.
Figure 3: Amino-acid sequence and northern expression of DRiP78 and its in vitro association with D1ct.
Figure 4: Yeast two-hybrid assay of D1ct with DRiP78.
Figure 5: Localization of DRiP78 and its effect on transport of the D1 receptor.
Figure 6: D1 receptor ligand binding regulation by DRiP78.
Figure 7: Pulse–chase analysis of the dopamine D1 receptor.

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Acknowledgements

We thank M. von Zastrow for Flag-containing constructs and F. Ehlert for M2 receptor cDNA and radioligand. We also thank O. Civelli for discussions, Z. Wang and C. Li for efforts in the early phase, and S. Vhan for technical assistance. This work was supported by grants from the NIH and the Alfred E. Sloan Foundation, and by an Optical Biology Shared Resource Grant.

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  1. Department of Pharmacology, University of California, Irvine, 92697, California, USA

    Jason C. Bermak, Ming Li, Clayton Bullock & Qun-Yong Zhou

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Correspondence to Qun-Yong Zhou.

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Bermak, J., Li, M., Bullock, C. et al. Regulation of transport of the dopamine D1 receptor by a new membrane-associated ER protein. Nat Cell Biol 3, 492–498 (2001). https://doi.org/10.1038/35074561

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