TY - JOUR
T1 - Functional characterization of C21ORF2 association with the NEK1 kinase mutated in human in diseases
AU - Gregorczyk, Mateusz
AU - Pastore, Graziana
AU - Muñoz, Ivan
AU - Carroll, Thomas
AU - Streubel, Johanna
AU - Munro, Meagan
AU - Lis, Pawel
AU - Lange, Sven
AU - Lamoliatte, Frederic
AU - Macartney, Thomas
AU - Toth, Rachel
AU - Brown, Fiona
AU - Hastie, James
AU - Pereira, Gislene
AU - Durocher, Daniel
AU - Rouse, John
N1 - Funding Information:
This work was supported by the Medical Research Council (grant number MC_UU_12016/1; M Gregorczyk, P Lis, F Lamoliatte, I Muñoz, J Rouse), by a grant from the Canadian Institutes for Health Research (grant PJT180438 to D Durocher) and by Merck K Ga A (Florian Weil and J Rouse).
Copyright:
© 2023 Gregorczyk et al.
PY - 2023/7
Y1 - 2023/7
N2 - The NEK1 kinase controls ciliogenesis, mitosis, and DNA repair, and NEK1 mutations cause human diseases including axial spondylometaphyseal dysplasia and amyotrophic lateral sclerosis. C21ORF2 mutations cause a similar pattern of human diseases, suggesting close functional links with NEK1 Here, we report that endogenous NEK1 and C21ORF2 form a tight complex in human cells. A C21ORF2 interaction domain "CID" at the C-terminus of NEK1 is necessary for its association with C21ORF2 in cells, and pathogenic mutations in this region disrupt the complex. AlphaFold modelling predicts an extended binding interface between a leucine-rich repeat domain in C21ORF2 and the NEK1-CID, and our model may explain why pathogenic mutations perturb the complex. We show that NEK1 mutations that inhibit kinase activity or weaken its association with C21ORF2 severely compromise ciliogenesis, and that C21ORF2, like NEK1 is required for homologous recombination. These data enhance our understanding of how the NEK1 kinase is regulated, and they shed light on NEK1-C21ORF2-associated diseases.
AB - The NEK1 kinase controls ciliogenesis, mitosis, and DNA repair, and NEK1 mutations cause human diseases including axial spondylometaphyseal dysplasia and amyotrophic lateral sclerosis. C21ORF2 mutations cause a similar pattern of human diseases, suggesting close functional links with NEK1 Here, we report that endogenous NEK1 and C21ORF2 form a tight complex in human cells. A C21ORF2 interaction domain "CID" at the C-terminus of NEK1 is necessary for its association with C21ORF2 in cells, and pathogenic mutations in this region disrupt the complex. AlphaFold modelling predicts an extended binding interface between a leucine-rich repeat domain in C21ORF2 and the NEK1-CID, and our model may explain why pathogenic mutations perturb the complex. We show that NEK1 mutations that inhibit kinase activity or weaken its association with C21ORF2 severely compromise ciliogenesis, and that C21ORF2, like NEK1 is required for homologous recombination. These data enhance our understanding of how the NEK1 kinase is regulated, and they shed light on NEK1-C21ORF2-associated diseases.
KW - Humans
KW - NIMA-Related Kinase 1/genetics
KW - Mutation/genetics
KW - DNA Repair
KW - Osteochondrodysplasias/genetics
KW - Phosphorylation
UR - http://www.scopus.com/inward/record.url?scp=85159419082&partnerID=8YFLogxK
U2 - 10.26508/lsa.202201740
DO - 10.26508/lsa.202201740
M3 - Article
C2 - 37188479
SN - 2575-1077
VL - 6
JO - Life Science Alliance
JF - Life Science Alliance
IS - 7
M1 - e202201740
ER -
