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Molecular Cloning of Mouse Homologue of Enamel Protein C4orf26 and Its Phosphorylation by FAM20C

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Abstract

It is widely accepted that cellular processes are controlled by protein phosphorylation and has become increasingly clear that protein degradation, localization and conformation as well as protein–protein interaction are the examples of subsequent cellular events modulated by protein phosphorylation. Enamel matrix proteins belong to members of the secretory calcium binding phosphoprotein (SCPP) family clustered on chromosome 4q21, and most of the SCPP phosphoproteins have at least one S-X-E motifs (S; serine, X; any amino acid, E; glutamic acid). It has been reported that mutations in C4orf26 gene, located on chromosome 4q21, are associated with autosomal recessive type of Amelogenesis Imperfecta (AI), a hereditary condition that affects enamel formation/mineralization. The enamel phenotype observed in patients with C4orf26 mutations is hypomineralized and partially hypoplastic, indicating that C4orf26 protein may function at both secretory and maturation stages of amelogenesis. The previous in vitro study showed that the synthetic phosphorylated peptide based on C4orf26 protein sequence accelerates hydroxyapatite nucleation. Here we show the molecular cloning of Gm1045, mouse homologue of C4orf26, which has 2 splicing isoforms. Immunohistochemical analysis demonstrated that the immunolocalization of Gm1045 is mainly observed in enamel matrix in vivo. Our report is the first to show that FAM20C, the Golgi casein kinase, phosphorylates C4orf26 and Gm1045 in cell cultures. The extracellular localization of C4orf26/Gm1045 was regulated by FAM20C kinase activity. Thus, our data point out the biological importance of enamel matrix-kinase control of SCPP phosphoproteins and may have a broad impact on the regulation of amelogenesis and AI.

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Acknowledgements

We thank Dr. Hidemitsu Harada (Iwate Medical University, Japan) for providing HAT-7 cells. We also thank for Dr. Sundharamani Venkitapathi for his assistance. This study was supported by grants from the National Institute of Dental and Craniofacial Research, NIH (DE019527 to YM.) and Boston University Henry Goldman School of Dental Medicine (YM.). NG and PP were supported by The Chulalongkorn Academic Advancement Into Its 2nd Century Project.

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Authors and Affiliations

  1. Department of Molecular and Cell Biology, Henry M. Goldman School of Dental Medicine, Boston University, Boston, MA, 02118, USA

    Nattanan Govitvattana, Yoshio Ohyama, Haytham Jaha, I-Ping Lin, Hanna Mochida & Yoshiyuki Mochida

  2. Department of Pediatric Dentistry, Faculty of Dentistry, Chulalongkorn University, Bangkok, 10330, Thailand

    Nattanan Govitvattana

  3. Division of Bio-Prosthodontics, Faculty of Dentistry & Graduate School of Medical and Dental Sciences, Niigata University, Niigata, 951-8514, Japan

    Masaru Kaku

  4. Graduate Institute of Clinical Dentistry, School of Dentistry, College of Medicine, National Taiwan University, Taipei, 10048, Taiwan

    I-Ping Lin

  5. Center of Excellence in Regenerative Dentistry, Faculty of Dentistry, Chulalongkorn University, Bangkok, 10330, Thailand

    Prasit Pavasant

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  1. Nattanan Govitvattana
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Contributions

Conceived and designed the experiments: NG, MK and YM. Performed the experiments: NG, MK, YO, HJ and IPL. Analyzed the data: NG, MK, PP, HM and YM. Wrote the paper: NG, MK, HM, PP and YM. Read and approved the final manuscript by all authors.

Corresponding authors

Correspondence to Prasit Pavasant or Yoshiyuki Mochida.

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Conflict of interest

Nattanan Govitvattana, Masaru Kaku, Yoshio Ohyama, Haytham Jaha, I-Ping Lin, Hanna Mochida, Prasit Pavasant and Yoshiyuki Mochida have declares no competing financial interests.

Ethical approval

The use of animals and all animal procedures in this study were approved by the Institutional Animal Care and Use Committee (IACUC) of Boston University Medical campus (approved protocol number: AN-15053), and all efforts were made to minimize suffering animals. This study was carried out in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health.

Accession Number

The nucleotide sequences for mouse Gm1045-a and Gm1045-b genes have been deposited in the GenBank database under GenBank Accession numbers; KF477194 and KC602377, respectively.

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Govitvattana, N., Kaku, M., Ohyama, Y. et al. Molecular Cloning of Mouse Homologue of Enamel Protein C4orf26 and Its Phosphorylation by FAM20C. Calcif Tissue Int 109, 445–454 (2021). https://doi.org/10.1007/s00223-021-00847-y

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