Abstract
Autosomal recessive microcephaly and chorioretinopathy (MCCRP) is a neurodevelopmental disorder characterized by delayed psychomotor development, growth retardation with dwarfism, and ocular abnormalities, and its occurrence has been found to be closely related to variants of the gene encoding centrosomes. However, the association between centrosomal duplication defects and the etiology of microcephaly syndromes is poorly understood. It is well known that polo-like kinase 4 (PLK4) is a key regulator of centriole duplication, and the abnormalities of centrosomal function caused by its protein variation need to be further explored in the pathogenesis of microcephaly. In our study, we found that a patient with microcephaly and chorioretinopathy harbored compound heterozygous missense variants NM_014264.4: c.2221C > T (p.Gln741*) and NM_014264.4: c.2062 T > C (p.Tyr688His) in the PLK4 gene. Overexpression experiments of the variant PLK4 proteins then showed that the G741 variant rather than the T688H variant had lost centrosomal amplification ability, and the G741 variant but not the T688H variant induced centrosomal replication disorder, which further inhibited cell proliferation, cycle division and cytoskeleton morphology in HeLa cells. Moreover, the overexpression of the two variant proteins had inconsistent effects on the target protein PLK4 by western blot analysis, also indicating that T688H variant overexpression is not functionally equivalent to WT-PLK4 overexpression. Therefore, all data support the idea that the PLK4 mutation induces centriolar duplication disorder and reduces the efficiency of mitosis inducing cell death or cell proliferation in the etiology of microcephaly disorder.
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Abbreviations
- MCCRP:
-
Autosomal recessive microcephaly and chorioretinopathy
- MCPH:
-
Autosomal recessive microcephaly
- PLK4:
-
Polo-like kinase 4
- MTT:
-
3-[4,5-Dimethyl-2-thiazolyl]-2,5-diphenyl-2H-tetrazolium bromide
- siRNA:
-
Small interfering RNA
- FBS:
-
Fetal bovine serum
- DMEM:
-
Dulbecco's modified Eagle's medium
- PMSF:
-
Phenylmethanesulfonyl fluoride
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Acknowledgements
The authors gratefully acknowledge the financial support from the Xiamen Municipal Medical and Health Guidance Project (3502Z20209200).
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The authors gratefully acknowledge the financial support from the Xiamen Municipal Medical and Health Guidance Project (3502Z20209200).
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Qi-Chang Wu, Ya-Song Xu, Zhi-Ying Su, Yi-Zhen Ji and Su-Qing Zhang conceived and performed the study. Li Sun, Xiao-Mei Yang, Shi-Yu Sun and Xiao-Fei Ji analyzed data. Ya-Song Xu and Zhi-Ying Su wrote the paper. Jie Tian and Qi-Chang Wu reviewed and edited the manuscript. All authors read and approved the manuscript.
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The genetic testing used in this study was approved by the ethical committees of Xiamen Maternal and Child Health Hospital in accordance with the Declaration of Helsinki. All patients provided informed consent, and all protocols were approved by the institutional committee of Xiamen Maternal and Child Health Hospital.
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Xu, YS., Su, ZY., Sun, L. et al. Overexpression of the PLK4 Gene as a Novel Strategy for the Treatment of Autosomal Recessive Microcephaly by Improving Centrosomal Dysfunction. J Mol Neurosci 71, 2618–2627 (2021). https://doi.org/10.1007/s12031-021-01881-z
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DOI: https://doi.org/10.1007/s12031-021-01881-z