Alternative splicing of the mouse amelogenin primary RNA transcript contributes to amelogenin heterogeneity

Lau, Eduardo C.; Simmer, James P.; Bringas, Pablo; Hsu, Dora D-J.; Hu, Ching-Chun; Zeichner‐David, Margarita; Thiemann, Flávia Torreão; Snead, Malcolm L.; Slavkin, Harold C.; Fincham, Alan G.

doi:10.1016/0006-291x(92)91366-x

Cited by 111 publications

(72 citation statements)

References 32 publications

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“…In mice, nine gene products of amelogenin have been identified and detected as protein, all of which contain exon 7 (5,6,14) in addition to the recently identified exon 8/9 (36). In rat, the amelogenin mRNAs that have been identified, with the exception of the exon 7 types, are R195 (contains exons 2, 3, 4, 5, 6ABCD, and 7), R181 (exons 2, 3, 5, 6ABCD, and 7), R73 (exons 2, 3, 4, 5, 6D, and 7), R59 (exons 2, 3, 5, 6D, and 7) (13), and R2-156 (exons 2, 3, 5 6BCD, and 7) (10).…”

Section: Discussionmentioning

confidence: 99%

The Exon 6ABC Region of Amelogenin mRNA Contribute to Increased Levels of Amelogenin mRNA through Amelogenin Protein-enhanced mRNA Stabilization

Harada

Taniguchi

2006

Journal of Biological Chemistry

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We recently demonstrated that the reuptake of full-length amelogenin protein results in increased levels of amelogenin mRNA through enhanced mRNA stabilization (Xu, L., Harada, H., Tamaki, T. Y., Matsumoto, S., Tanaka, J., and Taniguchi, A. (2006) J. Biol. Chem. 281, 2257-2262). Here, we examined the molecular mechanism of enhanced amelogenin mRNA stabilization. To identify the cis-regulatory region within amelogenin mRNA, we tested various reporter systems using a deletion series of reporter plasmids. A deletion at exon 6ABC of amelogenin mRNA resulted in a 2.5-fold increase in the amelogenin mRNA expression level when compared with that of full-length mRNA, indicating that a cis-element exists in exon 6ABC of amelogenin mRNA. Furthermore, Northwestern analysis demonstrated that amelogenin protein binds directly to its mRNA in vitro, suggesting that amelogenin protein acts as a trans-acting protein that specifically binds to this cis-element. Moreover, recombinant mouse amelogenin protein extended the half-life of full-length amelogenin mRNA but did not significantly alter the half-life of exon 6ABC-deletion mutant mRNA. The splice products produced by deletion of exon 6ABC are known as leucine-rich amelogenin peptides and have signaling effects on cells. Our findings also suggest that the regulation of full-length amelogenin protein expression differs from the regulation of leucine-rich amelogenin peptide expression.Amelogenin is a major component of enamel matrix. Amelogenin is unique in its localization on both X-chromosomes and Y-chromosomes in cows, pigs, and humans and on the X-chromosomes in mice (1-4) rather than on chromosome 4q like other enamel-and mineralization-associated proteins. Alternative splicing of amelogenin pre-mRNA leads to the production of many isoforms (5-11). The smaller splice products, produced upon the deletion of exon 6ABC, are known as the leucine-rich amelogenin peptides or LRAPs.2 Both full-length amelogenin and LRAP are capable of modulating expression of cementoblast-associated genes (12-16). In particular, LRAPs have been shown to act differently, as signaling molecules affecting odontogenic and other cell types (12-15). The larger forms, those that contain the intact proline-rich, hydrophobic exon 6 domains, are important for enamel mineralization (for review, see Ref. 17). Thus, the mRNA products of short and full-length amelogenin have different functions, and the regulation of full-length amelogenin protein expression appears to differ from the regulation of LRAP expression. CCAAT/enhancer-binding protein ␣ plays a key role in the developmentally regulated expression of the amelogenin gene at the transcription level (18), whereas Msx2 mediates interference with the binding of CCAAT/enhancer-binding protein ␣ to its cognate site on the mouse amelogenin minimal promoter by protein-protein interaction (19). Our previous study indicated that the reuptake of full-length amelogenin protein results in increased levels of amelogenin mRNA through enhanced mRNA stabilization (20)....

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Section: Discussionmentioning

confidence: 99%

The Exon 6ABC Region of Amelogenin mRNA Contribute to Increased Levels of Amelogenin mRNA through Amelogenin Protein-enhanced mRNA Stabilization

Harada

Taniguchi

2006

Journal of Biological Chemistry

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“…1) were used: (i) rM179 (20.16 kDa), a recombinant mouse amelogenin, which is identical to the native murine amelogenin, M180 (except for the lack of the amino-terminal methionine residue (10) and a phosphorylated serine at position 16) (11,12); (ii) rM166 (18.6 kDa), as rM179 but lacking the 13 C-terminal amino acid residues (13); (iii) TRAP (5.20 kDa), a synthetic murine tyrosine-rich amelogenin polypeptide representing the N-terminal 45 amino acid residues of the M180 amelogenin; (iv) LRAP (6.82 kDa), synthetic leucine-rich amelogenin polypeptide, identical to the full-length (M180) amelogenin at its two termini but lacking the center portion of the protein (14); (v) amelogenin C-terminal peptide (ACP); (vi) P173 (25 kDa) and P148 (20 kDa) (porcine amelogenins were extracted and purified following the protocol of Fincham et al (15) as described previously (5)); and (vii) ATMP, PYPSYGYEPMGGW and two altered ATMP peptides in one of which 1 The abbreviations used are: TRAP, tyrosine-rich amelogenin polypeptide; LRAP, leucine-rich amelogenin polypeptide; WGA, wheat germ agglutinin; GMp, GlcNAc mimicking peptide; ATMP, amelogenin trityrosyl motif peptide; T-ATMP, ATMP where proline is replaced by threonine; F-ATMP, ATMP where tyrosine is substituted by phenylalanine; ACP, amelogenin carboxyl-terminal peptide; HA, hemagglutination; HAI, hemagglutination inhibition; HSA, human serum albumin; HPLC, high performance liquid chromatography; TBS, Tris-buffered saline; AI, amelogenesis imperfecta; GM-peptide, GlcNAc-mimicking peptide; CK-14, cytokeratin-14; PVDF, polyvinylidene difluoride; BSA, bovine serum albumin. the third proline is substituted with threonine (T-ATMP) (PYPSYG-YETMGGW), and in another, all three tyrosine residues are replaced by phenylalanine (F-ATMP) (PFPSFGFEPMGGW) as described earlier (5) (see Fig.…”

Section: Methodsmentioning

confidence: 99%

The Enamel Protein Amelogenin Binds to the N-Acetyl-d-glucosamine-mimicking Peptide Motif of Cytokeratins

Ravindranath

Tam

Nguyen

et al. 2000

Journal of Biological Chemistry

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“…Several alternatively spliced isoforms of amelogenin mRNA (5,6), and their corresponding translation products (7), have been identified in the ameloblasts of developing mouse molars. The mouse gene contains 9 exons (8), but all potential splice isoforms are not equally expressed.…”

Section: Introductionmentioning

confidence: 99%

Identification of the functional activity of the [A-4] amelogenin gene splice product in newborn mouse ameloblasts

Iacob¹,

Veis²

2008

Bone

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In the mouse tooth organ, shortly after birth, ameloblasts acquire their secretory phenotype, which is characterized by the prominent expression and subsequent secretion of two isoforms of amelogenin, M180 and M59 (LRAP, [A−4]). Amelogenin deposition into the ameloblast extracellular matrix promotes enamel biomineralization. A complex set of intercellular signaling events, reciprocal communications between the developing oral epithelium and its underlying dental mesenchyme, guide the expression of amelogenin mRNA, and limit it to a defined period of tooth development. In tooth germ organ culture, addition of the [A−4] isoform, lacking amelogenin exon 4 and exon 6 segments a, b, c, was shown to affect ameloblast development. To understand the basis for this regulatory activity, we have studied the effects of r

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Alternative splicing of the mouse amelogenin primary RNA transcript contributes to amelogenin heterogeneity

Cited by 111 publications

References 32 publications

The Exon 6ABC Region of Amelogenin mRNA Contribute to Increased Levels of Amelogenin mRNA through Amelogenin Protein-enhanced mRNA Stabilization

The Exon 6ABC Region of Amelogenin mRNA Contribute to Increased Levels of Amelogenin mRNA through Amelogenin Protein-enhanced mRNA Stabilization

The Enamel Protein Amelogenin Binds to the N-Acetyl-d-glucosamine-mimicking Peptide Motif of Cytokeratins

Identification of the functional activity of the [A-4] amelogenin gene splice product in newborn mouse ameloblasts

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