Ma, P, Yan, W, Tian, Y et al. (3 more authors) (2016) The Importance of Serine Phosphorylation of Ameloblastin on Enamel Formation. Journal of Dental Research, 95 (12). pp. 1408-1414. ISSN 0022-0345
Abstract
FAM20C is a newly identified kinase on the secretory pathway responsible for the phosphorylation of serine residues in the Ser-x-Glu/pSer motifs in several enamel matrix proteins. Fam20C-knockout mice showed severe enamel defects very similar to those in the ameloblastin (Ambn)–knockout mice, implying that phosphoserines may have a critical role in AMBN function. To test this hypothesis, we generated amelogenin (Amel) promoter-driven Ambn-transgenic mice, in which Ser⁴⁸, Ser²²⁶, and Ser²²⁷ were replaced by aspartic acid (designated as D-Tg) or alanines (designated as A-Tg). The negative charge of aspartic acid is believed to be able to mimic the phosphorylation state of serine, while alanine is a commonly used residue to substitute serine due to their similar structure. Using Western immunoblotting and quantitative polymerase chain reaction, the authors identified transgenic lines expressing transgenes somewhat higher (Tg+) or much higher (Tg++) than endogenous Ambn. The lower incisors collected from 7-d-old and 7-wk-old mice were analyzed by histology, scanning electron microscopy, immunohistochemistry, and Western immunoblotting to examine the morphology and microstructure changes in enamel, as well as the expression pattern of enamel matrix proteins. The A-Tg+ and A-Tg++ mice displayed severe enamel defects in spite of the expression level of transgenes, while the D-Tg+ and D-Tg++ mice showed minor to mild enamel defects, indicating that the D-Tg transgenes disturbed enamel formation less than the A-Tg transgenes did. Our results suggest that the phosphorylation state of serines is likely an essential component for the integrity of AMBN function.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | (c) 2016, International & American Associations for Dental Research. This is an author produced version of a paper published in the Journal of Dental Research. Uploaded in accordance with the publisher's self-archiving policy. |
Keywords: | phosphoserine; FAM20C; amelogenesis; transgene; extracellular matrix; kinase |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Medicine and Health (Leeds) > School of Dentistry (Leeds) > Oral Biology (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 07 Oct 2016 09:43 |
Last Modified: | 27 Mar 2020 01:27 |
Published Version: | https://doi.org/10.1177/0022034516661513 |
Status: | Published |
Publisher: | SAGE Publications |
Identification Number: | 10.1177/0022034516661513 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:105538 |