Alternative splicing regulates adaptor protein binding, trafficking, and activity of the Vps10p domain receptor SorCS2 in neuronal development

Sune Skeldal; Lasse Frank Voss; Jonas Lende; Sarah Broholt Pedersen; Simon Mølgaard; Mathias Kaas; Perline Demange; Andreas Høiberg Bentsen; Marie Fuglsang; Marie Rubin Sander; Henriette Buttenschøn; Camilla Gustafsen; Peder Madsen; Simon Glerup

doi:10.1016/j.jbc.2023.105102

Alternative splicing regulates adaptor protein binding, trafficking, and activity of the Vps10p domain receptor SorCS2 in neuronal development

Sune Skeldal, Lasse Frank Voss, Jonas Lende, Sarah Broholt Pedersen, Simon Mølgaard, Mathias Kaas, Perline Demange, Andreas Høiberg Bentsen, Marie Fuglsang, Marie Rubin Sander, Henriette Buttenschøn, Camilla Gustafsen, Peder Madsen^*, Simon Glerup^*

^*Corresponding author for this work

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

Abstract

The Vps10p domain receptor SorCS2 is crucial for the development and function of the nervous system and essential for brain-derived neurotrophic factor (BDNF)-induced changes in neuronal morphology and plasticity. SorCS2 regulates the subcellular trafficking of the BDNF signaling receptor TrkB as well as selected neurotransmitter receptors in a manner that is dependent on the SorCS2 intracellular domain (ICD). However, the cellular machinery and adaptor protein (AP) interactions that regulate receptor trafficking via the SorCS2 ICD are unknown. We here identify four splice variants of human SorCS2 differing in the insertion of an acidic cluster motif and/or a serine residue within the ICD. We show that each variant undergoes posttranslational proteolytic processing into a one- or two-chain receptor, giving rise to eight protein isoforms, the expression of which differs between neuronal and nonneuronal tissues and is affected by cellular stressors. We found that the only variants without the serine were able to rescue BDNF-induced branching of SorCS2 knockout hippocampal neurons, while variants without the acidic cluster showed increased interactions with clathrin-associated APs AP-1, AP-2, and AP-3. Using yeast two-hybrid screens, we further discovered that all variants bound dynein light chain Tctex-type 3; however, only variants with an acidic cluster motif bound kinesin light chain 1. Accordingly, splice variants showed markedly different trafficking properties and localized to different subcellular compartments. Taken together, our findings demonstrate the existence of eight functional SorCS2 isoforms with differential capacity for interactions with cytosolic ligands dynein light chain Tctex-type 3 and kinesin light chain 1, which potentially allows cell-type specific SorCS2 trafficking and BDNF signaling.

Original language	English
Article number	105102
Journal	Journal of Biological Chemistry
Volume	299
Issue	9
Number of pages	15
ISSN	0021-9258
DOIs	https://doi.org/10.1016/j.jbc.2023.105102
Publication status	Published - Sept 2023

Keywords

alternative splicing
BDNF
cellular stress
dynein
kinesin
receptor trafficking
SorCS2

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Skeldal, S., Voss, L. F., Lende, J., Pedersen, S. B., Mølgaard, S., Kaas, M., Demange, P., Bentsen, A. H., Fuglsang, M., Sander, M. R., Buttenschøn, H., Gustafsen, C., Madsen, P., & Glerup, S. (2023). Alternative splicing regulates adaptor protein binding, trafficking, and activity of the Vps10p domain receptor SorCS2 in neuronal development. Journal of Biological Chemistry, 299(9), Article 105102. https://doi.org/10.1016/j.jbc.2023.105102

@article{fdb3f461baec458f81895fa93f273a41,

title = "Alternative splicing regulates adaptor protein binding, trafficking, and activity of the Vps10p domain receptor SorCS2 in neuronal development",

abstract = "The Vps10p domain receptor SorCS2 is crucial for the development and function of the nervous system and essential for brain-derived neurotrophic factor (BDNF)-induced changes in neuronal morphology and plasticity. SorCS2 regulates the subcellular trafficking of the BDNF signaling receptor TrkB as well as selected neurotransmitter receptors in a manner that is dependent on the SorCS2 intracellular domain (ICD). However, the cellular machinery and adaptor protein (AP) interactions that regulate receptor trafficking via the SorCS2 ICD are unknown. We here identify four splice variants of human SorCS2 differing in the insertion of an acidic cluster motif and/or a serine residue within the ICD. We show that each variant undergoes posttranslational proteolytic processing into a one- or two-chain receptor, giving rise to eight protein isoforms, the expression of which differs between neuronal and nonneuronal tissues and is affected by cellular stressors. We found that the only variants without the serine were able to rescue BDNF-induced branching of SorCS2 knockout hippocampal neurons, while variants without the acidic cluster showed increased interactions with clathrin-associated APs AP-1, AP-2, and AP-3. Using yeast two-hybrid screens, we further discovered that all variants bound dynein light chain Tctex-type 3; however, only variants with an acidic cluster motif bound kinesin light chain 1. Accordingly, splice variants showed markedly different trafficking properties and localized to different subcellular compartments. Taken together, our findings demonstrate the existence of eight functional SorCS2 isoforms with differential capacity for interactions with cytosolic ligands dynein light chain Tctex-type 3 and kinesin light chain 1, which potentially allows cell-type specific SorCS2 trafficking and BDNF signaling.",

keywords = "alternative splicing, BDNF, cellular stress, dynein, kinesin, receptor trafficking, SorCS2",

author = "Sune Skeldal and Voss, {Lasse Frank} and Jonas Lende and Pedersen, {Sarah Broholt} and Simon M{\o}lgaard and Mathias Kaas and Perline Demange and Bentsen, {Andreas H{\o}iberg} and Marie Fuglsang and Sander, {Marie Rubin} and Henriette Buttensch{\o}n and Camilla Gustafsen and Peder Madsen and Simon Glerup",

year = "2023",

month = sep,

doi = "10.1016/j.jbc.2023.105102",

language = "English",

volume = "299",

journal = "Journal of Biological Chemistry",

issn = "0021-9258",

publisher = "American Society for Biochemistry and Molecular Biology, Inc.",

number = "9",

}

Skeldal, S, Voss, LF, Lende, J, Pedersen, SB , Mølgaard, S, Kaas, M, Demange, P, Bentsen, AH, Fuglsang, M, Sander, MR, Buttenschøn, H , Gustafsen, C , Madsen, P & Glerup, S 2023, 'Alternative splicing regulates adaptor protein binding, trafficking, and activity of the Vps10p domain receptor SorCS2 in neuronal development', Journal of Biological Chemistry, vol. 299, no. 9, 105102. https://doi.org/10.1016/j.jbc.2023.105102

Alternative splicing regulates adaptor protein binding, trafficking, and activity of the Vps10p domain receptor SorCS2 in neuronal development. / Skeldal, Sune; Voss, Lasse Frank; Lende, Jonas et al.
In: Journal of Biological Chemistry, Vol. 299, No. 9, 105102, 09.2023.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

TY - JOUR

T1 - Alternative splicing regulates adaptor protein binding, trafficking, and activity of the Vps10p domain receptor SorCS2 in neuronal development

AU - Skeldal, Sune

AU - Voss, Lasse Frank

AU - Lende, Jonas

AU - Pedersen, Sarah Broholt

AU - Mølgaard, Simon

AU - Kaas, Mathias

AU - Demange, Perline

AU - Bentsen, Andreas Høiberg

AU - Fuglsang, Marie

AU - Sander, Marie Rubin

AU - Buttenschøn, Henriette

AU - Gustafsen, Camilla

AU - Madsen, Peder

AU - Glerup, Simon

PY - 2023/9

Y1 - 2023/9

N2 - The Vps10p domain receptor SorCS2 is crucial for the development and function of the nervous system and essential for brain-derived neurotrophic factor (BDNF)-induced changes in neuronal morphology and plasticity. SorCS2 regulates the subcellular trafficking of the BDNF signaling receptor TrkB as well as selected neurotransmitter receptors in a manner that is dependent on the SorCS2 intracellular domain (ICD). However, the cellular machinery and adaptor protein (AP) interactions that regulate receptor trafficking via the SorCS2 ICD are unknown. We here identify four splice variants of human SorCS2 differing in the insertion of an acidic cluster motif and/or a serine residue within the ICD. We show that each variant undergoes posttranslational proteolytic processing into a one- or two-chain receptor, giving rise to eight protein isoforms, the expression of which differs between neuronal and nonneuronal tissues and is affected by cellular stressors. We found that the only variants without the serine were able to rescue BDNF-induced branching of SorCS2 knockout hippocampal neurons, while variants without the acidic cluster showed increased interactions with clathrin-associated APs AP-1, AP-2, and AP-3. Using yeast two-hybrid screens, we further discovered that all variants bound dynein light chain Tctex-type 3; however, only variants with an acidic cluster motif bound kinesin light chain 1. Accordingly, splice variants showed markedly different trafficking properties and localized to different subcellular compartments. Taken together, our findings demonstrate the existence of eight functional SorCS2 isoforms with differential capacity for interactions with cytosolic ligands dynein light chain Tctex-type 3 and kinesin light chain 1, which potentially allows cell-type specific SorCS2 trafficking and BDNF signaling.

AB - The Vps10p domain receptor SorCS2 is crucial for the development and function of the nervous system and essential for brain-derived neurotrophic factor (BDNF)-induced changes in neuronal morphology and plasticity. SorCS2 regulates the subcellular trafficking of the BDNF signaling receptor TrkB as well as selected neurotransmitter receptors in a manner that is dependent on the SorCS2 intracellular domain (ICD). However, the cellular machinery and adaptor protein (AP) interactions that regulate receptor trafficking via the SorCS2 ICD are unknown. We here identify four splice variants of human SorCS2 differing in the insertion of an acidic cluster motif and/or a serine residue within the ICD. We show that each variant undergoes posttranslational proteolytic processing into a one- or two-chain receptor, giving rise to eight protein isoforms, the expression of which differs between neuronal and nonneuronal tissues and is affected by cellular stressors. We found that the only variants without the serine were able to rescue BDNF-induced branching of SorCS2 knockout hippocampal neurons, while variants without the acidic cluster showed increased interactions with clathrin-associated APs AP-1, AP-2, and AP-3. Using yeast two-hybrid screens, we further discovered that all variants bound dynein light chain Tctex-type 3; however, only variants with an acidic cluster motif bound kinesin light chain 1. Accordingly, splice variants showed markedly different trafficking properties and localized to different subcellular compartments. Taken together, our findings demonstrate the existence of eight functional SorCS2 isoforms with differential capacity for interactions with cytosolic ligands dynein light chain Tctex-type 3 and kinesin light chain 1, which potentially allows cell-type specific SorCS2 trafficking and BDNF signaling.

KW - alternative splicing

KW - BDNF

KW - cellular stress

KW - dynein

KW - kinesin

KW - receptor trafficking

KW - SorCS2

UR - http://www.scopus.com/inward/record.url?scp=85168744159&partnerID=8YFLogxK

U2 - 10.1016/j.jbc.2023.105102

DO - 10.1016/j.jbc.2023.105102

M3 - Journal article

C2 - 37507021

AN - SCOPUS:85168744159

SN - 0021-9258

VL - 299

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 9

M1 - 105102

ER -

Alternative splicing regulates adaptor protein binding, trafficking, and activity of the Vps10p domain receptor SorCS2 in neuronal development

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