Abstract
The activity of transcription factors is often regulated by Post-translational modifications. A precondition for such modifications is the presence, in the corresponding mRNAs, of the exons that either directly encode the modifiable residues in question, or encode protein domains that influence their modification indirectly. The inclusion or exclusion of coding exons is regulated predominantly by alternative splicing but can also depend on promoter choice and polyadenylation site selection. Information about exon inclusion and exclusion, both qualitatively and quantitatively, is particularly important for experiments designed to mutate endogenous codons because such mutations can alter splicing patterns. Therefore, we here describe methods employed to quantitate exon inclusion and exclusion, using as example a mouse transcription factor gene, Mitf.
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References
Hodgkinson CA, Moore KJ, Nakayama A, Steingrimsson E, Copeland NG, Jenkins NA, Arnheiter H (1993) Mutations at the mouse microphthalmia locus are associated with defects in a gene encoding a novel basic-helix-loop-helix-zipper protein. Cell 74:395–404
Arnheiter H, Hou L, Nguyen MTT, Bismuth K, Csermely T, Murakami H, Skuntz S, Liu W, Bharti K (2006) Mitf – A matter of life and death for the developing melanocyte. In: Hearing V, Leong SPL (eds) From melanocytes to malignant melanoma. Humana, Totowa, NJ
Tassabehji M, Newton VE, Read AP (1994) Waardenburg syndrome type 2 caused by mutations in the human microphthalmia (MITF) gene. Nat Genet 8:251–255
Bharti K, Nguyen MT, Skuntz S, Bertuzzi S, Arnheiter H (2006) The other pigment cell: specification and development of the pigmented epithelium of the vertebrate eye. Pigment Cell Res 19:380–394
Steingrimsson E, Copeland NG, Jenkins NA (2004) Melanocytes and the microphthalmia transcription factor network. Annu Rev Genet 38:365–411
Bharti K, Liu W, Csermely T, Bertuzzi S, Arnheiter H (2008) Alternative promoter use in eye development: the complex role and regulation of the transcription factor MITF. Development 135:1169–1178
Hallsson JH, Favor J, Hodgkinson C, Glaser T, Lamoreux ML, Magnusdottir R, Gunnarsson GJ, Sweet HO, Copeland NG, Jenkins NA, Steingrimsson E (2000) Genomic, transcriptional and mutational analysis of the mouse microphthalmia locus. Genetics 155:291–300
Murakami H, Arnheiter H (2005) Sumoylation modulates transcriptional activity of MITF in a promoter-specific manner. Pigment Cell Res 18:265–277
Miller AJ, Levy C, Davis IJ, Razin E, Fisher DE (2005) Sumoylation of MITF and its related family members TFE3 and TFEB. J Biol Chem 280:146–155
Hemesath TJ, Price ER, Takemoto C, Badalian T, Fisher DE (1998) MAP kinase links the transcription factor Microphthalmia to c-Kit signalling in melanocytes. Nature 391:298–301
Wu M, Hemesath TJ, Takemoto CM, Horstmann MA, Wells AG, Price ER, Fisher DZ, Fisher DE (2000) c-Kit triggers dual phosphorylations, which couple activation and degradation of the essential melanocyte factor Mi. Genes Dev 14:301–312
Bismuth K, Skuntz S, Hallsson JH, Pak E, Dutra AS, Steingrimsson E, Arnheiter H (2008) An unstable targeted allele of the mouse Mitf gene with a high somatic and germline reversion rate. Genetics 178:259–272
Lin S, Fu XD (2007) SR proteins and related factors in alternative splicing. Adv Exp Med Biol 623:107–122
Nagy A, Gertenstein M, Vintersten K, Behringer R (2003) Manipulating the mouse embryo. A laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
Acknowledgments
This work was supported by the Intramural Research Program of the National Institutes of Health, NINDS.
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Bharti, K., Debbache, J., Wang, X., Arnheiter, H. (2010). The Basic-Helix-Loop-Helix-Leucine Zipper Gene Mitf: Analysis of Alternative Promoter Choice and Splicing. In: Higgins, P. (eds) Transcription Factors. Methods in Molecular Biology, vol 647. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-738-9_14
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DOI: https://doi.org/10.1007/978-1-60761-738-9_14
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