Expression and Transcriptional Activity of Alternative Splice Variants of Mitf Exon 6.
From: Laboratory of Molecular Biology, Azabu University School of Veterinary Medicine, Sagamihara, Japan.
Molecular and cellular biochemistry
- Publish Date: Sep 2007
- ISSN: 0300-8177
- Volume: 303
- Issue: 1-2
- Pages: 251-7
- Medium: Print
- Language: English
- Citation (JAMA): Murakami Masaru, Iwata Yasuhiro, Funaba Masayuki, et al. Expression and Transcriptional Activity of Alternative Splice Variants of Mitf Exon 6.. Mol. Cell. Biochem. Sep 2007;303:251-7
Abstract
Microphthalmia-associated transcription factor (Mitf) is a tissue-specific transcription factor. At least nine distinct mouse isoform mRNAs are encoded by alternative splicing of the first exon of Mitf (Mitf-A, -B, -C, -D, -E, -H, -J, -M, and -mc), while exons 2-9 of all Mitf isoforms examined to date are identical. In addition, alternative splice variants of exon 6a encoding 6 amino acid proximal to the basic region of the protein are known in Mitf-A, -H, and -M. In this study, we identified alternative splice variants of exon 6a in other Mitf isoforms (Mitf-E, -J, and -mc) in melanocytes, mast cells, macrophages, and heart. We also compared the transcriptional activity of Mitf variants containing exon 6a to that of Mitf variants that did not contain exon 6a. PCR-RFLP analysis revealed that expression of Mitf with exon 6a was comparable with that of Mitf without exon 6a, irrespective of the specificity of the first exon, or cell type, although Mitf isoforms with different first exons were expressed in a cell type-dependent manner. Luciferase-based reporter assays revealed that transcription of Tyrosinase, which is known Mitf-regulated gene, was elicited more efficiently by expression of Mitf isoforms containing exon 6a, compared to isoforms that did not contain exon 6a. However, when transcription of Tyrp-1, Mmcp-6, and PAI-1 was examined, no significant differences were detected between Mitf isoforms with exon 6a and those without exon 6a, except for Tyrp-1 transcription by Mitf-D/E isoform. These results reveal a diverse pattern of gene expression and different transcriptional activities of Mitf isoforms, suggesting discrete regulation of gene transcription in specific tissues by Mitf.
Mesh Headings (Keywords): Alternative Splicing, Animals, Cells, Cultured, Exons, Gene Expression Regulation, Heart, Humans, Macrophages, Mast Cells, Melanocytes, Melanoma, Experimental, Membrane Glycoproteins, Mice, Microphthalmia-Associated Transcription Factor, Monophenol Monooxygenase, Oxidoreductases, Plasminogen Activator Inhibitor 1, Polymerase Chain Reaction, Polymorphism, Restriction Fragment Length, Protein Isoforms, RNA, Messenger, Trans-Activation (Genetics), Transcription, Genetic, Tryptases
Check for Full Text / PubMed Unique Identifier (PMID): 17457519
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