CNKSR2 interactome analysis indicates its association with the centrosome/microtubule system

The protein connector enhancer of kinase suppressor of Ras 2 (CNKSR2), present in both the postsynaptic density and cytoplasm of neurons, is a scaffolding protein with several protein-binding domains. Variants of the CNKSR2 gene have been implicated in neurodevelopmental disorders, particularly inte...

Full description

Saved in:
Bibliographic Details
Main Authors: Lin Yin, Yalan Xu, Jie Mu, Yu Leng, Lei Ma, Yu Zheng, Ruizhi Li, Yin Wang, Peifeng Li, Hai Zhu, Dong Wang, Jing Li
Format: Article
Language:English
Published: Wolters Kluwer Medknow Publications 2025-08-01
Series:Neural Regeneration Research
Subjects:
Online Access:https://journals.lww.com/10.4103/NRR.NRR-D-23-01725
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850180852511670272
author Lin Yin
Yalan Xu
Jie Mu
Yu Leng
Lei Ma
Yu Zheng
Ruizhi Li
Yin Wang
Peifeng Li
Hai Zhu
Dong Wang
Jing Li
author_facet Lin Yin
Yalan Xu
Jie Mu
Yu Leng
Lei Ma
Yu Zheng
Ruizhi Li
Yin Wang
Peifeng Li
Hai Zhu
Dong Wang
Jing Li
author_sort Lin Yin
collection DOAJ
description The protein connector enhancer of kinase suppressor of Ras 2 (CNKSR2), present in both the postsynaptic density and cytoplasm of neurons, is a scaffolding protein with several protein-binding domains. Variants of the CNKSR2 gene have been implicated in neurodevelopmental disorders, particularly intellectual disability, although the precise mechanism involved has not yet been fully understood. Research has demonstrated that CNKSR2 plays a role in facilitating the localization of postsynaptic density protein complexes to the membrane, thereby influencing synaptic signaling and the morphogenesis of dendritic spines. However, the function of CNKSR2 in the cytoplasm remains to be elucidated. In this study, we used immunoprecipitation and high-resolution liquid chromatography-mass spectrometry to identify the interactors of CNKSR2. Through a combination of bioinformatic analysis and cytological experiments, we found that the CNKSR2 interactors were significantly enriched in the proteome of the centrosome. We also showed that CNKSR2 interacted with the microtubule protein DYNC1H1 and with the centrosome marker CEP290. Subsequent colocalization analysis confirmed the centrosomal localization of CNKSR2. When we downregulated CNKSR2 expression in mouse neuroblastoma cells (Neuro 2A), we observed significant changes in the expression of numerous centrosomal genes. This manipulation also affected centrosome-related functions, including cell size and shape, cell proliferation, and motility. Furthermore, we found that CNKSR2 interactors were highly enriched in de novo variants associated with intellectual disability and autism spectrum disorder. Our findings establish a connection between CNKSR2 and the centrosome, and offer new insights into the underlying mechanisms of neurodevelopmental disorders.
format Article
id doaj-art-e985cc5239864277b6f568fa3fc94495
institution OA Journals
issn 1673-5374
1876-7958
language English
publishDate 2025-08-01
publisher Wolters Kluwer Medknow Publications
record_format Article
series Neural Regeneration Research
spelling doaj-art-e985cc5239864277b6f568fa3fc944952025-08-20T02:18:01ZengWolters Kluwer Medknow PublicationsNeural Regeneration Research1673-53741876-79582025-08-012082420243210.4103/NRR.NRR-D-23-01725CNKSR2 interactome analysis indicates its association with the centrosome/microtubule systemLin YinYalan XuJie MuYu LengLei MaYu ZhengRuizhi LiYin WangPeifeng LiHai ZhuDong WangJing LiThe protein connector enhancer of kinase suppressor of Ras 2 (CNKSR2), present in both the postsynaptic density and cytoplasm of neurons, is a scaffolding protein with several protein-binding domains. Variants of the CNKSR2 gene have been implicated in neurodevelopmental disorders, particularly intellectual disability, although the precise mechanism involved has not yet been fully understood. Research has demonstrated that CNKSR2 plays a role in facilitating the localization of postsynaptic density protein complexes to the membrane, thereby influencing synaptic signaling and the morphogenesis of dendritic spines. However, the function of CNKSR2 in the cytoplasm remains to be elucidated. In this study, we used immunoprecipitation and high-resolution liquid chromatography-mass spectrometry to identify the interactors of CNKSR2. Through a combination of bioinformatic analysis and cytological experiments, we found that the CNKSR2 interactors were significantly enriched in the proteome of the centrosome. We also showed that CNKSR2 interacted with the microtubule protein DYNC1H1 and with the centrosome marker CEP290. Subsequent colocalization analysis confirmed the centrosomal localization of CNKSR2. When we downregulated CNKSR2 expression in mouse neuroblastoma cells (Neuro 2A), we observed significant changes in the expression of numerous centrosomal genes. This manipulation also affected centrosome-related functions, including cell size and shape, cell proliferation, and motility. Furthermore, we found that CNKSR2 interactors were highly enriched in de novo variants associated with intellectual disability and autism spectrum disorder. Our findings establish a connection between CNKSR2 and the centrosome, and offer new insights into the underlying mechanisms of neurodevelopmental disorders.https://journals.lww.com/10.4103/NRR.NRR-D-23-01725autism spectrum disordercentrosomecnksr2intellectual disabilityinteractomemass spectrometrymicrotubuleneurodevelopmental diseaseprotein complexesprotein–protein interactions
spellingShingle Lin Yin
Yalan Xu
Jie Mu
Yu Leng
Lei Ma
Yu Zheng
Ruizhi Li
Yin Wang
Peifeng Li
Hai Zhu
Dong Wang
Jing Li
CNKSR2 interactome analysis indicates its association with the centrosome/microtubule system
Neural Regeneration Research
autism spectrum disorder
centrosome
cnksr2
intellectual disability
interactome
mass spectrometry
microtubule
neurodevelopmental disease
protein complexes
protein–protein interactions
title CNKSR2 interactome analysis indicates its association with the centrosome/microtubule system
title_full CNKSR2 interactome analysis indicates its association with the centrosome/microtubule system
title_fullStr CNKSR2 interactome analysis indicates its association with the centrosome/microtubule system
title_full_unstemmed CNKSR2 interactome analysis indicates its association with the centrosome/microtubule system
title_short CNKSR2 interactome analysis indicates its association with the centrosome/microtubule system
title_sort cnksr2 interactome analysis indicates its association with the centrosome microtubule system
topic autism spectrum disorder
centrosome
cnksr2
intellectual disability
interactome
mass spectrometry
microtubule
neurodevelopmental disease
protein complexes
protein–protein interactions
url https://journals.lww.com/10.4103/NRR.NRR-D-23-01725
work_keys_str_mv AT linyin cnksr2interactomeanalysisindicatesitsassociationwiththecentrosomemicrotubulesystem
AT yalanxu cnksr2interactomeanalysisindicatesitsassociationwiththecentrosomemicrotubulesystem
AT jiemu cnksr2interactomeanalysisindicatesitsassociationwiththecentrosomemicrotubulesystem
AT yuleng cnksr2interactomeanalysisindicatesitsassociationwiththecentrosomemicrotubulesystem
AT leima cnksr2interactomeanalysisindicatesitsassociationwiththecentrosomemicrotubulesystem
AT yuzheng cnksr2interactomeanalysisindicatesitsassociationwiththecentrosomemicrotubulesystem
AT ruizhili cnksr2interactomeanalysisindicatesitsassociationwiththecentrosomemicrotubulesystem
AT yinwang cnksr2interactomeanalysisindicatesitsassociationwiththecentrosomemicrotubulesystem
AT peifengli cnksr2interactomeanalysisindicatesitsassociationwiththecentrosomemicrotubulesystem
AT haizhu cnksr2interactomeanalysisindicatesitsassociationwiththecentrosomemicrotubulesystem
AT dongwang cnksr2interactomeanalysisindicatesitsassociationwiththecentrosomemicrotubulesystem
AT jingli cnksr2interactomeanalysisindicatesitsassociationwiththecentrosomemicrotubulesystem