CNKSR2 Interactome Analysis Indicates Its Association with the Centrosome/microtubule System

Overview

Journal Neural Regen Res

Date 2024 Oct 3

PMID 39359098

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

Affiliations

Soon will be listed here.

Abstract

JOURNAL/nrgr/04.03/01300535-202508000-00031/figure1/v/2024-09-30T120553Z/r/image-tiff 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.

References

Akhmanova A, Kapitein L . Mechanisms of microtubule organization in differentiated animal cells. Nat Rev Mol Cell Biol. 2022; 23(8):541-558. DOI: 10.1038/s41580-022-00473-y. View

. Diverse genetic causes of polymicrogyria with epilepsy. Epilepsia. 2021; 62(4):973-983. PMC: 10838185. DOI: 10.1111/epi.16854. View

Loffler H, Fechter A, Matuszewska M, Saffrich R, Mistrik M, Marhold J . Cep63 recruits Cdk1 to the centrosome: implications for regulation of mitotic entry, centrosome amplification, and genome maintenance. Cancer Res. 2011; 71(6):2129-39. DOI: 10.1158/0008-5472.CAN-10-2684. View

Vaags A, Bowdin S, Smith M, Gilbert-Dussardier B, Brocke-Holmefjord K, Sinopoli K . Absent CNKSR2 causes seizures and intellectual, attention, and language deficits. Ann Neurol. 2014; 76(5):758-64. DOI: 10.1002/ana.24274. View

Nigg E, Raff J . Centrioles, centrosomes, and cilia in health and disease. Cell. 2009; 139(4):663-78. DOI: 10.1016/j.cell.2009.10.036. View

Li J, Wilkinson B, Clementel V, Hou J, ODell T, Coba M . Long-term potentiation modulates synaptic phosphorylation networks and reshapes the structure of the postsynaptic interactome. Sci Signal. 2016; 9(440):rs8. DOI: 10.1126/scisignal.aaf6716. View

Allen A, Cossette P, Delanty N, Eichler E, Goldstein D, Han Y . De novo mutations in epileptic encephalopathies. Nature. 2013; 501(7466):217-21. PMC: 3773011. DOI: 10.1038/nature12439. View

Meka D, Scharrenberg R, de Anda F . Emerging roles of the centrosome in neuronal development. Cytoskeleton (Hoboken). 2020; 77(3-4):84-96. DOI: 10.1002/cm.21593. View

Lim E, Uddin M, De Rubeis S, Chan Y, Kamumbu A, Zhang X . Rates, distribution and implications of postzygotic mosaic mutations in autism spectrum disorder. Nat Neurosci. 2017; 20(9):1217-1224. PMC: 5672813. DOI: 10.1038/nn.4598. View

10.

Xu Y, Mu J, Zhou Z, Leng Y, Yu Y, Song X . Expansion of mouse castration-resistant intermediate prostate stem cells in vitro. Stem Cell Res Ther. 2022; 13(1):299. PMC: 9284701. DOI: 10.1186/s13287-022-02978-x. View

11.

Sun Y, Liu Y, Xu Z, Kong Q, Wang Y . mutation causes the X-linked epilepsy-aphasia syndrome: A case report and review of literature. World J Clin Cases. 2018; 6(12):570-576. PMC: 6212609. DOI: 10.12998/wjcc.v6.i12.570. View

12.

Zieger H, Kunde S, Rademacher N, Schmerl B, Shoichet S . Disease-associated synaptic scaffold protein CNK2 modulates PSD size and influences localisation of the regulatory kinase TNIK. Sci Rep. 2020; 10(1):5709. PMC: 7109135. DOI: 10.1038/s41598-020-62207-4. View

13.

Griff J, Langlie J, Bencie N, Cromar Z, Mittal J, Memis I . Recent advancements in noninvasive brain modulation for individuals with autism spectrum disorder. Neural Regen Res. 2022; 18(6):1191-1195. PMC: 9838164. DOI: 10.4103/1673-5374.360163. View

14.

Wilfert A, Sulovari A, Turner T, Coe B, Eichler E . Recurrent de novo mutations in neurodevelopmental disorders: properties and clinical implications. Genome Med. 2017; 9(1):101. PMC: 5704398. DOI: 10.1186/s13073-017-0498-x. View

15.

Lanigan T, Liu A, Huang Y, Mei L, Margolis B, Guan K . Human homologue of Drosophila CNK interacts with Ras effector proteins Raf and Rlf. FASEB J. 2003; 17(14):2048-60. DOI: 10.1096/fj.02-1096com. View

16.

Fromer M, Pocklington A, Kavanagh D, Williams H, Dwyer S, Gormley P . De novo mutations in schizophrenia implicate synaptic networks. Nature. 2014; 506(7487):179-84. PMC: 4237002. DOI: 10.1038/nature12929. View

17.

Kang Q, Yang L, Liao H, Wu L, Chen B, Yang S . CNKSR2 gene mutation leads to Houge type of X-linked syndromic mental retardation: A case report and review of literature. Medicine (Baltimore). 2021; 100(23):e26093. PMC: 8202604. DOI: 10.1097/MD.0000000000026093. View

18.

Pinto D, Delaby E, Merico D, Barbosa M, Merikangas A, Klei L . Convergence of genes and cellular pathways dysregulated in autism spectrum disorders. Am J Hum Genet. 2014; 94(5):677-94. PMC: 4067558. DOI: 10.1016/j.ajhg.2014.03.018. View

19.

Li K, Savitska D, Garaschuk O . K+ channel-mediated retarded maturation of interneurons and its role in neurodevelopmental disorders. Neural Regen Res. 2023; 19(7):1403-1404. PMC: 10883523. DOI: 10.4103/1673-5374.386409. View

20.

Di Gioia S, Letteboer S, Kostic C, Bandah-Rozenfeld D, Hetterschijt L, Sharon D . FAM161A, associated with retinitis pigmentosa, is a component of the cilia-basal body complex and interacts with proteins involved in ciliopathies. Hum Mol Genet. 2012; 21(23):5174-84. DOI: 10.1093/hmg/dds368. View