The DEP Domain Determines Subcellular Targeting of the GTPase Activating Protein RGS9 In Vivo

Kirill A. Martemyanov, Polina V. Lishko, Nidia Calero, Gabor Keresztes, Maxim Sokolov, Katherine J. Strissel, Ilya B. Leskov, Johnathan A. Hopp, Alexander V. Kolesnikov, Ching Kang Chen, Janis Lem, Stefan Heller, Marie E Burns, Vadim Y. Arshavsky

Research output: Contribution to journalArticle

98 Citations (Scopus)

Abstract

DEP (for Disheveled, EGL-10, Pleckstrin) homology domains are present in numerous signaling proteins, including many in the nervous system, but their function remains mostly elusive. We report that the DEP domain of a photoreceptor-specific signaling protein, RGS9 (for regulator of G-protein signaling 9), plays an essential role in RGS9 delivery to the intracellular compartment of its functioning, the rod outer segment. We generated a transgenic mouse in which RGS9 was replaced by its mutant lacking the DEP domain. We then used a combination of the quantitative technique of serial tangential sectioning-Western blotting with electrophysiological recordings to demonstrate that mutant RGS9 is expressed in rods in the normal amount but is completely excluded from the outer segments. The delivery of RGS9 to rod outer segments is likely to be mediated by the DEP domain interaction with a transmembrane protein, R9AP (for RGS9 anchoring protein), known to anchor RGS9 on the surface of photoreceptor membranes and to potentiate RGS9 catalytic activity. We show that both of these functions are also abolished as the result of the DEP domain deletion. These findings indicate that a novel function of the DEP domain is to target a signaling protein to a specific compartment of a highly polarized neuron. Interestingly, sequence analysis of R9AP reveals the presence of a conserved R-SNARE (for soluble N-ethylmaleimide-sensitive factor attachment protein receptor) motif and a predicted overall structural homology with SNARE proteins involved in vesicular trafficking and fusion. This presents the possibility that DEP domains might serve to target various DEP-containing proteins to the sites of their intracellular action via interactions with the members of extended SNARE protein family.

Original languageEnglish (US)
Pages (from-to)10175-10181
Number of pages7
JournalJournal of Neuroscience
Volume23
Issue number32
StatePublished - Nov 12 2003

Fingerprint

GTP-Binding Protein Regulators
GTPase-Activating Proteins
SNARE Proteins
RGS Proteins
Rod Cell Outer Segment
Proteins
Amino Acid Motifs
platelet protein P47
Nervous System
Transgenic Mice
Sequence Analysis
Western Blotting
Neurons
Membranes

Keywords

  • DEP domains
  • Photoreceptors
  • Protein targeting
  • Retina
  • RGS proteins
  • SNARE proteins

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Martemyanov, K. A., Lishko, P. V., Calero, N., Keresztes, G., Sokolov, M., Strissel, K. J., ... Arshavsky, V. Y. (2003). The DEP Domain Determines Subcellular Targeting of the GTPase Activating Protein RGS9 In Vivo. Journal of Neuroscience, 23(32), 10175-10181.

The DEP Domain Determines Subcellular Targeting of the GTPase Activating Protein RGS9 In Vivo. / Martemyanov, Kirill A.; Lishko, Polina V.; Calero, Nidia; Keresztes, Gabor; Sokolov, Maxim; Strissel, Katherine J.; Leskov, Ilya B.; Hopp, Johnathan A.; Kolesnikov, Alexander V.; Chen, Ching Kang; Lem, Janis; Heller, Stefan; Burns, Marie E; Arshavsky, Vadim Y.

In: Journal of Neuroscience, Vol. 23, No. 32, 12.11.2003, p. 10175-10181.

Research output: Contribution to journalArticle

Martemyanov, KA, Lishko, PV, Calero, N, Keresztes, G, Sokolov, M, Strissel, KJ, Leskov, IB, Hopp, JA, Kolesnikov, AV, Chen, CK, Lem, J, Heller, S, Burns, ME & Arshavsky, VY 2003, 'The DEP Domain Determines Subcellular Targeting of the GTPase Activating Protein RGS9 In Vivo', Journal of Neuroscience, vol. 23, no. 32, pp. 10175-10181.
Martemyanov KA, Lishko PV, Calero N, Keresztes G, Sokolov M, Strissel KJ et al. The DEP Domain Determines Subcellular Targeting of the GTPase Activating Protein RGS9 In Vivo. Journal of Neuroscience. 2003 Nov 12;23(32):10175-10181.
Martemyanov, Kirill A. ; Lishko, Polina V. ; Calero, Nidia ; Keresztes, Gabor ; Sokolov, Maxim ; Strissel, Katherine J. ; Leskov, Ilya B. ; Hopp, Johnathan A. ; Kolesnikov, Alexander V. ; Chen, Ching Kang ; Lem, Janis ; Heller, Stefan ; Burns, Marie E ; Arshavsky, Vadim Y. / The DEP Domain Determines Subcellular Targeting of the GTPase Activating Protein RGS9 In Vivo. In: Journal of Neuroscience. 2003 ; Vol. 23, No. 32. pp. 10175-10181.
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AU - Sokolov, Maxim

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AU - Kolesnikov, Alexander V.

AU - Chen, Ching Kang

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