Neuronal ER-plasma Membrane Junctions Organized by Kv2-VAP Pairing Recruit Nir Proteins and Affect Phosphoinositide Homeostasis

Overview

Journal J Biol Chem

Specialty Biochemistry

Date 2019 Oct 10

PMID 31594866

Citations 20

Authors

Michael Kirmiz

Taryn E Gillies

Eamonn J Dickson

James S Trimmer

Affiliations

Soon will be listed here.

Abstract

The association of plasma membrane (PM)-localized voltage-gated potassium (Kv2) channels with endoplasmic reticulum (ER)-localized vesicle-associated membrane protein-associated proteins VAPA and VAPB defines ER-PM junctions in mammalian brain neurons. Here, we used proteomics to identify proteins associated with Kv2/VAP-containing ER-PM junctions. We found that the VAP-interacting membrane-associated phosphatidylinositol (PtdIns) transfer proteins PYK2 N-terminal domain-interacting receptor 2 (Nir2) and Nir3 specifically associate with Kv2.1 complexes. When coexpressed with Kv2.1 and VAPA in HEK293T cells, Nir2 colocalized with cell-surface-conducting and -nonconducting Kv2.1 isoforms. This was enhanced by muscarinic-mediated PtdIns(4,5)P hydrolysis, leading to dynamic recruitment of Nir2 to Kv2.1 clusters. In cultured rat hippocampal neurons, exogenously expressed Nir2 did not strongly colocalize with Kv2.1, unless exogenous VAPA was also expressed, supporting the notion that VAPA mediates the spatial association of Kv2.1 and Nir2. Immunolabeling signals of endogenous Kv2.1, Nir2, and VAP puncta were spatially correlated in cultured neurons. Fluorescence-recovery-after-photobleaching experiments revealed that Kv2.1, VAPA, and Nir2 have comparable turnover rates at ER-PM junctions, suggesting that they form complexes at these sites. Exogenous Kv2.1 expression in HEK293T cells resulted in significant differences in the kinetics of PtdIns(4,5)P recovery following repetitive muscarinic stimulation, with no apparent impact on resting PtdIns(4,5)P or PtdIns(4)P levels. Finally, the brains of Kv2.1-knockout mice had altered composition of PtdIns lipids, suggesting a crucial role for native Kv2.1-containing ER-PM junctions in regulating PtdIns lipid metabolism in brain neurons. These results suggest that ER-PM junctions formed by Kv2 channel-VAP pairing regulate PtdIns lipid homeostasis via VAP-associated PtdIns transfer proteins.

Citing Articles

Mammalian START-like phosphatidylinositol transfer proteins - Physiological perspectives and roles in cancer biology.

Pathak A, Willis K, Bankaitis V, McDermott M Biochim Biophys Acta Mol Cell Biol Lipids. 2024; 1869(7):159529.

PMID: 38945251 PMC: 11533902. DOI: 10.1016/j.bbalip.2024.159529.

Downregulation of the silent potassium channel Kv8.1 increases motor neuron vulnerability in amyotrophic lateral sclerosis.

Huang X, Lee S, Chen K, Kawaguchi R, Wiskow O, Ghosh S Brain Commun. 2024; 6(3):fcae202.

PMID: 38911266 PMC: 11191651. DOI: 10.1093/braincomms/fcae202.

The tether function of the anoctamins.

Lin W, Chung W, Muallem S Cell Calcium. 2024; 121:102875.

PMID: 38701708 PMC: 11166512. DOI: 10.1016/j.ceca.2024.102875.

Outlining cardiac ion channel protein interactors and their signature in the human electrocardiogram.

Maurya S, Mills R, Kahnert K, Chiang D, Bertoli G, Lundegaard P Nat Cardiovasc Res. 2024; 2(7):673-692.

PMID: 38666184 PMC: 11041666. DOI: 10.1038/s44161-023-00294-y.

Electrically silent KvS subunits associate with native Kv2 channels in brain and impact diverse properties of channel function.

Ferns M, van der List D, Vierra N, Lacey T, Murray K, Kirmiz M bioRxiv. 2024; .

PMID: 38328147 PMC: 10849721. DOI: 10.1101/2024.01.25.577135.

References

Herculano-Houzel S, Mota B, Lent R . Cellular scaling rules for rodent brains. Proc Natl Acad Sci U S A. 2006; 103(32):12138-43. PMC: 1567708. DOI: 10.1073/pnas.0604911103. View

Kirmiz M, Palacio S, Thapa P, King A, Sack J, Trimmer J . Remodeling neuronal ER-PM junctions is a conserved nonconducting function of Kv2 plasma membrane ion channels. Mol Biol Cell. 2018; 29(20):2410-2432. PMC: 6233057. DOI: 10.1091/mbc.E18-05-0337. View

OConnell K, Rolig A, Whitesell J, Tamkun M . Kv2.1 potassium channels are retained within dynamic cell surface microdomains that are defined by a perimeter fence. J Neurosci. 2006; 26(38):9609-18. PMC: 6674455. DOI: 10.1523/JNEUROSCI.1825-06.2006. View

Kim Y, Guzman-Hernandez M, Wisniewski E, Balla T . Phosphatidylinositol-Phosphatidic Acid Exchange by Nir2 at ER-PM Contact Sites Maintains Phosphoinositide Signaling Competence. Dev Cell. 2015; 33(5):549-61. PMC: 4476625. DOI: 10.1016/j.devcel.2015.04.028. View

Murakoshi H, Trimmer J . Identification of the Kv2.1 K+ channel as a major component of the delayed rectifier K+ current in rat hippocampal neurons. J Neurosci. 1999; 19(5):1728-35. PMC: 6782166. View

Takeshima H, Komazaki S, Nishi M, Iino M, Kangawa K . Junctophilins: a novel family of junctional membrane complex proteins. Mol Cell. 2000; 6(1):11-22. DOI: 10.1016/s1097-2765(00)00003-4. View

Henne W, Liou J, Emr S . Molecular mechanisms of inter-organelle ER-PM contact sites. Curr Opin Cell Biol. 2015; 35:123-30. DOI: 10.1016/j.ceb.2015.05.001. View

Lee H, Wang J, Swartz K . Interaction between extracellular Hanatoxin and the resting conformation of the voltage-sensor paddle in Kv channels. Neuron. 2003; 40(3):527-36. DOI: 10.1016/s0896-6273(03)00636-6. View

Tao-Cheng J . Activity-dependent decrease in contact areas between subsurface cisterns and plasma membrane of hippocampal neurons. Mol Brain. 2018; 11(1):23. PMC: 5902880. DOI: 10.1186/s13041-018-0366-7. View

10.

McKenzie A, Wang M, Hauberg M, Fullard J, Kozlenkov A, Keenan A . Brain Cell Type Specific Gene Expression and Co-expression Network Architectures. Sci Rep. 2018; 8(1):8868. PMC: 5995803. DOI: 10.1038/s41598-018-27293-5. View

11.

Nishimura Y, Hayashi M, Inada H, Tanaka T . Molecular cloning and characterization of mammalian homologues of vesicle-associated membrane protein-associated (VAMP-associated) proteins. Biochem Biophys Res Commun. 1999; 254(1):21-6. DOI: 10.1006/bbrc.1998.9876. View

12.

Du J, Tao-Cheng J, Zerfas P, McBain C . The K+ channel, Kv2.1, is apposed to astrocytic processes and is associated with inhibitory postsynaptic membranes in hippocampal and cortical principal neurons and inhibitory interneurons. Neuroscience. 1998; 84(1):37-48. DOI: 10.1016/s0306-4522(97)00519-8. View

13.

Kruse M, Vivas O, Traynor-Kaplan A, Hille B . Dynamics of Phosphoinositide-Dependent Signaling in Sympathetic Neurons. J Neurosci. 2016; 36(4):1386-400. PMC: 4728732. DOI: 10.1523/JNEUROSCI.3535-15.2016. View

14.

Lev S, Ben Halevy D, Peretti D, Dahan N . The VAP protein family: from cellular functions to motor neuron disease. Trends Cell Biol. 2008; 18(6):282-90. DOI: 10.1016/j.tcb.2008.03.006. View

15.

Sun E, Guillen-Samander A, Bian X, Wu Y, Cai Y, Messa M . Lipid transporter TMEM24/C2CD2L is a Ca-regulated component of ER-plasma membrane contacts in mammalian neurons. Proc Natl Acad Sci U S A. 2019; 116(12):5775-5784. PMC: 6431226. DOI: 10.1073/pnas.1820156116. View

16.

Henkart M, Landis D, Reese T . Similarity of junctions between plasma membranes and endoplasmic reticulum in muscle and neurons. J Cell Biol. 1976; 70(2 pt 1):338-47. PMC: 2109828. DOI: 10.1083/jcb.70.2.338. View

17.

Wu Y, Whiteus C, Xu C, Hayworth K, Weinberg R, Hess H . Contacts between the endoplasmic reticulum and other membranes in neurons. Proc Natl Acad Sci U S A. 2017; 114(24):E4859-E4867. PMC: 5474793. DOI: 10.1073/pnas.1701078114. View

18.

Rocha N, Kuijl C, van der Kant R, Janssen L, Houben D, Janssen H . Cholesterol sensor ORP1L contacts the ER protein VAP to control Rab7-RILP-p150 Glued and late endosome positioning. J Cell Biol. 2009; 185(7):1209-25. PMC: 2712958. DOI: 10.1083/jcb.200811005. View

19.

Sarmiere P, Weigle C, Tamkun M . The Kv2.1 K+ channel targets to the axon initial segment of hippocampal and cortical neurons in culture and in situ. BMC Neurosci. 2008; 9:112. PMC: 2592246. DOI: 10.1186/1471-2202-9-112. View

20.

King A, Manning C, Trimmer J . A unique ion channel clustering domain on the axon initial segment of mammalian neurons. J Comp Neurol. 2014; 522(11):2594-608. PMC: 4133991. DOI: 10.1002/cne.23551. View