Fam83h is Associated with Intracellular Vesicles and ADHCAI

Abstract

Defects in FAM83H on human chromosome 8q24.3 cause autosomal-dominant hypocalcified amelogenesis imperfecta (ADHCAI). FAM83H does not encode a recognizable signal peptide, so we predicted that the Fam83h protein functions within the cell. We tested this hypothesis by constitutively expressing mouse Fam83h with green fluorescent protein (GFP) fused to its C-terminus in HEK293 and HeLa cell lines. Green fluorescent signal from the Fam83h-GFP fusion protein was associated with perinuclear vesicles, usually in the vicinity of the Golgi apparatus. No signal was observed within the nucleus. In addition, we identified FAM83H nonsense mutations in Hispanic (C1330C>T; p.Q444X) and Caucasian (c.1192C>T; p.Q398X) families with ADHCAI. We conclude that Fam83h localizes in the intracellular environment, is associated with vesicles, and plays an important role in dental enamel formation. FAM83H is the first gene involved in the etiology of amelogenesis imperfecta (AI) that does not encode a secreted protein.

Get full access to this article

View all access options for this article.

References

Bartlett JD, Simmer JP, Xue J, Margolis HC, Moreno EC (1996). Molecular cloning and mRNA tissue distribution of a novel matrix metalloproteinase isolated from porcine enamel organ. Gene 183:123–128.

Cerny R, Slaby I, Hammarström L, Wurtz T (1996). A novel gene expressed in rat ameloblasts codes for proteins with cell binding domains. J Bone Min Res 11:883–891.

Du G, Huang P, Liang BT, Frohman MA (2004). Phospholipase D2 localizes to the plasma membrane and regulates angiotensin II receptor endocytosis. Mol Biol Cell 15:1024–1030.

Hart PS, Becerik S, Cogulu D, Emingil G, Ozdemir-Ozenen D, Han ST, et al. (2009). Novel FAM83H mutations in Turkish families with auto-somal dominant hypocalcified amelogenesis imperfecta. Clin Genet 75:401–404.

Hiroyama M, Exton J (2005). Localization and regulation of phospholipase D2 by ARF6. J Cell Biochem 95:149–164.

Hu CC, Zhang C, Qian Q, Ryu OH, Moradian-Oldak J, Fincham AG, et al. (1996). Cloning, DNA sequence, and alternative splicing of opossum amelogenin mRNAs. J Dent Res 75:1728–1734.

Hu CC, Fukae M, Uchida T, Qian Q, Zhang CH, Ryu OH, et al. (1997a). Sheathlin: cloning, cDNA/polypeptide sequences, and immunolocalization of porcine enamel proteins concentrated in the sheath space. J Dent Res 76:648–657.

Hu CC, Fukae M, Uchida T, Qian Q, Zhang CH, Ryu OH, et al. (1997b). Cloning and characterization of porcine enamelin mRNAs. J Dent Res 76:1720–1729.

Hu CC, Simmer JP, Bartlett JD, Qian Q, Zhang C, Ryu OH, et al. (1998). Murine enamelin: cDNA and derived protein sequences. Connect Tissue Res 39:47–61.

10.

Hu JC, Chun YH, Al Hazzazzi T, Simmer JP (2007). Enamel formation and amelogenesis imperfecta. Cells Tissues Organs 186:78–85.

11.

Iwasaki K, Bajenova E, Somogyi-Ganss E, Miller M, Nguyen V, Nourkeyhani H, et al. (2005). Amelotin—a novel secreted, ameloblast-specific protein. J Dent Res 84:1127–1132.

12.

Kim JW, Simmer JP, Lin BP, Seymen F, Bartlett JD, Hu JC (2006). Mutational analysis of candidate genes in 24 amelogenesis imperfecta families. Eur J Oral Sci 114(Suppl 1):3–12.

13.

Kim JW, Lee SK, Lee ZH, Park JC, Lee KE, Lee MH, et al. (2008). FAM83H mutations in families with autosomal-dominant hypocalcified amelogenesis imperfecta. Am J Hum Genet 82:489–494.

14.

Krebsbach PH, Lee SK, Matsuki Y, Kozak CA, Yamada K, Yamada Y (1996). Full-length sequence, localization, and chromosomal mapping of ameloblastin: a novel tooth-specific gene. J Biol Chem 271:4431–4435.

15.

Lee SK, Hu JC, Bartlett JD, Lee KE, Lin BP, Simmer JP, et al. (2008). Mutational spectrum of FAM83H: the C-terminal portion is required for tooth enamel calcification. Hum Mutat 29:E95–E99.

16.

Marchler-Bauer A, Anderson J, Derbyshire M, DeWeese-Scott C, Gonzales N, Gwadz M, et al. (2007). CDD: a conserved domain database for interactive domain family analysis. Nucleic Acids Res 35:D237–D240.

17.

Mendoza G, Pemberton TJ, Lee K, Scarel-Caminaga R, Mehrian-Shai R, Gonzalez-Quevedo C, et al. (2007). A new locus for autosomal dominant amelogenesis imperfecta on chromosome 8q24.3. Hum Genet 120:653–662.

18.

Moffatt P, Smith CE, St-Arnaud R, Simmons D, Wright JT, Nanci A (2006). Cloning of rat amelotin and localization of the protein to the basal lamina of maturation stage ameloblasts and junctional epithelium. Biochem J 399:37–46.

19.

Moffatt P, Smith CE, St-Arnaud R, Nanci A (2008). Characterization of Apin, a secreted protein highly expressed in tooth-associated epithelia. J Cell Biochem 103:941–956.

20.

Shyu AB, Wilkinson MF, van Hoof A (2008). Messenger RNA regulation: to translate or to degrade. EMBO J 27:471–481.

21.

Simmer JP, Fukae M, Tanabe T, Yamakoshi Y, Uchida T, Xue J, et al. (1998). Purification, characterization, and cloning of enamel matrix serine proteinase 1. J Dent Res 77:377–386.

22.

Snead ML, Zeichner-David M, Chandra T, Robson KJ, Woo SL, Slavkin HC (1983). Construction and identification of mouse amelogenin cDNA clones. Proc Natl Acad Sci USA 80:7254–7258.

23.

Snead ML, Lau EC, Zeichner-David M, Fincham AG, Woo SL, Slavkin HC (1985). DNA sequence for cloned cDNA for murine amelogenin reveal the amino acid sequence for enamel-specific protein. Biochem Biophys Res Commun 129:812–818.

24.

Takagi T, Suzuki M, Baba T, Minegishi K, Sasaki S (1984). Complete amino acid sequence of amelogenin in developing bovine enamel. Biochem Biophys Res Commun 121:592–597.

25.

Witkop CJ Jr, Sauk JJ Jr (1976). Heritable defects of enamel. In: Oral facial genetics. Stewart RE, Prescott GH, editors. St. Louis: C.V. Mosby Co., pp. 151–226.

26.

Wright JT, Hart TC, Hart PS, Simmons D, Suggs C, Daley B, et al. (2009a). Human and mouse enamel phenotypes resulting from mutation or altered expression of AMEL, ENAM, MMP20 and KLK4. Cells Tissues Organs 189:224–229.

27.

Wright JT, Frazier-Bowers S, Simmons D, Alexander K, Crawford P, Han ST, et al. (2009b). Phenotypic variation in FAM83H-associated amelo-genesis imperfecta. J Dent Res 88:356–360.

Supplementary Material

Please find the following supplemental material available below.

For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.

For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.

0.00 MB

0.33 MB