Interactome network analysis identifies multiple caspase-6 interactors involved in the pathogenesis of HD

Sean-Patrick Riechers; Stefanie Butland; Yu Deng; Niels Skotte; Dagmar E Ehrnhoefer; Jenny Russ; Jean Laine; Melissa Laroche; Mahmoud A Pouladi; Erich E Wanker; Michael R Hayden; Rona K Graham

doi:10.1093/hmg/ddw036

Interactome network analysis identifies multiple caspase-6 interactors involved in the pathogenesis of HD

Sean-Patrick Riechers, Stefanie Butland, Yu Deng, Niels Skotte, Dagmar E Ehrnhoefer, Jenny Russ, Jean Laine, Melissa Laroche, Mahmoud A Pouladi, Erich E Wanker, Michael R Hayden, Rona K Graham

11 Citations (Scopus)

Abstract

Caspase-6 (CASP6) has emerged as an important player in Huntington disease (HD), Alzheimer disease (AD) and cerebral ischemia, where it is activated early in the disease process. CASP6 also plays a key role in axonal degeneration, further underscoring the importance of this protease in neurodegenerative pathways. As a protein's function is modulated by its protein-protein interactions, we performed a high-throughput yeast-2-hybrid (Y2H) screen against ∼17,000 human proteins to gain further insight into the function of CASP6. We identified a high-confidence list of 87 potential CASP6 interactors. From this list, 61% are predicted to contain a CASP6 recognition site. Of nine candidate substrates assessed, six are cleaved by CASP6. Proteins that did not contain a predicted CASP6 recognition site were assessed using a LUMIER assay approach, and 51% were further validated as interactors by this method. Of note, 54% of the high-confidence interactors identified show alterations in human HD brain at the mRNA level, and there is a significant enrichment for previously validated huntingtin (HTT) interactors. One protein of interest, STK3, a pro-apoptotic kinase, was validated biochemically to be a CASP6 substrate. Furthermore, our results demonstrate that in striatal cells expressing mutant huntingtin (mHTT), an increase in full length and fragment levels of STK3 are observed. We further show that caspase-3 is not essential for the endogenous cleavage of STK3. Characterization of the interaction network provides important new information regarding key pathways of interactors of CASP6 and highlights potential novel therapeutic targets for HD, AD and cerebral ischemia.

Original language	English
Journal	Human Molecular Genetics
Volume	25
Issue number	8
Pages (from-to)	1600-18
Number of pages	19
ISSN	0964-6906
DOIs	https://doi.org/10.1093/hmg/ddw036
Publication status	Published - 15 Apr 2016
Externally published	Yes

Keywords

Binding Sites
Caspase 6
Cell Line
Gene Expression Regulation
Humans
Huntingtin Protein
Huntington Disease
Models, Biological
Protein Interaction Maps
Protein Processing, Post-Translational
Protein-Serine-Threonine Kinases
Two-Hybrid System Techniques
Journal Article
Research Support, Non-U.S. Gov't

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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@article{81f310d3636b4347a8cd052b7b65e73b,

title = "Interactome network analysis identifies multiple caspase-6 interactors involved in the pathogenesis of HD",

abstract = "Caspase-6 (CASP6) has emerged as an important player in Huntington disease (HD), Alzheimer disease (AD) and cerebral ischemia, where it is activated early in the disease process. CASP6 also plays a key role in axonal degeneration, further underscoring the importance of this protease in neurodegenerative pathways. As a protein's function is modulated by its protein-protein interactions, we performed a high-throughput yeast-2-hybrid (Y2H) screen against ∼17,000 human proteins to gain further insight into the function of CASP6. We identified a high-confidence list of 87 potential CASP6 interactors. From this list, 61% are predicted to contain a CASP6 recognition site. Of nine candidate substrates assessed, six are cleaved by CASP6. Proteins that did not contain a predicted CASP6 recognition site were assessed using a LUMIER assay approach, and 51% were further validated as interactors by this method. Of note, 54% of the high-confidence interactors identified show alterations in human HD brain at the mRNA level, and there is a significant enrichment for previously validated huntingtin (HTT) interactors. One protein of interest, STK3, a pro-apoptotic kinase, was validated biochemically to be a CASP6 substrate. Furthermore, our results demonstrate that in striatal cells expressing mutant huntingtin (mHTT), an increase in full length and fragment levels of STK3 are observed. We further show that caspase-3 is not essential for the endogenous cleavage of STK3. Characterization of the interaction network provides important new information regarding key pathways of interactors of CASP6 and highlights potential novel therapeutic targets for HD, AD and cerebral ischemia.",

keywords = "Binding Sites, Caspase 6, Cell Line, Gene Expression Regulation, Humans, Huntingtin Protein, Huntington Disease, Models, Biological, Protein Interaction Maps, Protein Processing, Post-Translational, Protein-Serine-Threonine Kinases, Two-Hybrid System Techniques, Journal Article, Research Support, Non-U.S. Gov't",

author = "Sean-Patrick Riechers and Stefanie Butland and Yu Deng and Niels Skotte and Ehrnhoefer, {Dagmar E} and Jenny Russ and Jean Laine and Melissa Laroche and Pouladi, {Mahmoud A} and Wanker, {Erich E} and Hayden, {Michael R} and Graham, {Rona K}",

year = "2016",

month = apr,

day = "15",

doi = "10.1093/hmg/ddw036",

language = "English",

volume = "25",

pages = "1600--18",

journal = "Human Molecular Genetics",

issn = "0964-6906",

publisher = "Oxford University Press",

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T1 - Interactome network analysis identifies multiple caspase-6 interactors involved in the pathogenesis of HD

AU - Riechers, Sean-Patrick

AU - Butland, Stefanie

AU - Deng, Yu

AU - Skotte, Niels

AU - Ehrnhoefer, Dagmar E

AU - Russ, Jenny

AU - Laine, Jean

AU - Laroche, Melissa

AU - Pouladi, Mahmoud A

AU - Wanker, Erich E

AU - Hayden, Michael R

AU - Graham, Rona K

PY - 2016/4/15

Y1 - 2016/4/15

N2 - Caspase-6 (CASP6) has emerged as an important player in Huntington disease (HD), Alzheimer disease (AD) and cerebral ischemia, where it is activated early in the disease process. CASP6 also plays a key role in axonal degeneration, further underscoring the importance of this protease in neurodegenerative pathways. As a protein's function is modulated by its protein-protein interactions, we performed a high-throughput yeast-2-hybrid (Y2H) screen against ∼17,000 human proteins to gain further insight into the function of CASP6. We identified a high-confidence list of 87 potential CASP6 interactors. From this list, 61% are predicted to contain a CASP6 recognition site. Of nine candidate substrates assessed, six are cleaved by CASP6. Proteins that did not contain a predicted CASP6 recognition site were assessed using a LUMIER assay approach, and 51% were further validated as interactors by this method. Of note, 54% of the high-confidence interactors identified show alterations in human HD brain at the mRNA level, and there is a significant enrichment for previously validated huntingtin (HTT) interactors. One protein of interest, STK3, a pro-apoptotic kinase, was validated biochemically to be a CASP6 substrate. Furthermore, our results demonstrate that in striatal cells expressing mutant huntingtin (mHTT), an increase in full length and fragment levels of STK3 are observed. We further show that caspase-3 is not essential for the endogenous cleavage of STK3. Characterization of the interaction network provides important new information regarding key pathways of interactors of CASP6 and highlights potential novel therapeutic targets for HD, AD and cerebral ischemia.

AB - Caspase-6 (CASP6) has emerged as an important player in Huntington disease (HD), Alzheimer disease (AD) and cerebral ischemia, where it is activated early in the disease process. CASP6 also plays a key role in axonal degeneration, further underscoring the importance of this protease in neurodegenerative pathways. As a protein's function is modulated by its protein-protein interactions, we performed a high-throughput yeast-2-hybrid (Y2H) screen against ∼17,000 human proteins to gain further insight into the function of CASP6. We identified a high-confidence list of 87 potential CASP6 interactors. From this list, 61% are predicted to contain a CASP6 recognition site. Of nine candidate substrates assessed, six are cleaved by CASP6. Proteins that did not contain a predicted CASP6 recognition site were assessed using a LUMIER assay approach, and 51% were further validated as interactors by this method. Of note, 54% of the high-confidence interactors identified show alterations in human HD brain at the mRNA level, and there is a significant enrichment for previously validated huntingtin (HTT) interactors. One protein of interest, STK3, a pro-apoptotic kinase, was validated biochemically to be a CASP6 substrate. Furthermore, our results demonstrate that in striatal cells expressing mutant huntingtin (mHTT), an increase in full length and fragment levels of STK3 are observed. We further show that caspase-3 is not essential for the endogenous cleavage of STK3. Characterization of the interaction network provides important new information regarding key pathways of interactors of CASP6 and highlights potential novel therapeutic targets for HD, AD and cerebral ischemia.

KW - Binding Sites

KW - Caspase 6

KW - Cell Line

KW - Gene Expression Regulation

KW - Humans

KW - Huntingtin Protein

KW - Huntington Disease

KW - Models, Biological

KW - Protein Interaction Maps

KW - Protein Processing, Post-Translational

KW - Protein-Serine-Threonine Kinases

KW - Two-Hybrid System Techniques

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1093/hmg/ddw036

DO - 10.1093/hmg/ddw036

M3 - Journal article

C2 - 26908611

SN - 0964-6906

VL - 25

SP - 1600

EP - 1618

JO - Human Molecular Genetics

JF - Human Molecular Genetics

IS - 8

ER -

Interactome network analysis identifies multiple caspase-6 interactors involved in the pathogenesis of HD

Abstract

Keywords

UN SDGs

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