The solution structure of DNA-free Pax-8 paired box domain accounts for redox regulation of transcriptional activity in the pax protein family
- PMID: 18829450
- PMCID: PMC2662260
- DOI: 10.1074/jbc.M805717200
The solution structure of DNA-free Pax-8 paired box domain accounts for redox regulation of transcriptional activity in the pax protein family
Abstract
Pax-8 is a transcription factor belonging to the PAX genes superfamily and its crucial role has been proven both in embryo and in the adult organism. Pax-8 activity is regulated via a redox-based mechanism centered on the glutathionylation of specific cysteines in the N-terminal region (Cys45 and Cys57). These residues belong to a highly evolutionary conserved DNA binding site: the Paired Box (Prd) domain. Crystallographic protein-DNA complexes of the homologues Pax-6 and Pax-5 showed a bipartite Prd domain consisting of two helix-turn-helix (HTH) motifs separated by an extended linker region. Here, by means of nuclear magnetic resonance, we show for the first time that the HTH motifs are largely defined in the unbound Pax-8 Prd domain. Our findings contrast with previous induced fit models, in which Pax-8 is supposed to largely fold upon DNA binding. Importantly, our data provide the structural basis for the enhanced chemical reactivity of residues Cys45 and Cys57 and explain clinical missense mutations that are not obviously related to the DNA binding interface of the paired box domain. Finally, sequence conservation suggests that our findings could be a general feature of the Pax family transcription factors.
Figures






Similar articles
-
Gene regulation by PAX6: structural-functional correlations of missense mutants and transcriptional control of Trpm3/miR-204.Mol Vis. 2014 Mar 6;20:270-82. eCollection 2014. Mol Vis. 2014. PMID: 24623969 Free PMC article.
-
Lune/eye gone, a Pax-like protein, uses a partial paired domain and a homeodomain for DNA recognition.Proc Natl Acad Sci U S A. 1998 Nov 10;95(23):13720-5. doi: 10.1073/pnas.95.23.13720. Proc Natl Acad Sci U S A. 1998. PMID: 9811867 Free PMC article.
-
Crystal structure of the human Pax6 paired domain-DNA complex reveals specific roles for the linker region and carboxy-terminal subdomain in DNA binding.Genes Dev. 1999 May 15;13(10):1263-75. doi: 10.1101/gad.13.10.1263. Genes Dev. 1999. PMID: 10346815 Free PMC article.
-
PAX proteins and fables of their reconstruction.Crit Rev Eukaryot Gene Expr. 2012;22(2):161-77. doi: 10.1615/critreveukargeneexpr.v22.i2.70. Crit Rev Eukaryot Gene Expr. 2012. PMID: 22856433 Review.
-
Focus on molecules: Pax-6, the eye master.Exp Eye Res. 2006 Aug;83(2):233-4. doi: 10.1016/j.exer.2005.11.019. Epub 2006 Mar 23. Exp Eye Res. 2006. PMID: 16563385 Review. No abstract available.
Cited by
-
Overview of PAX gene family: analysis of human tissue-specific variant expression and involvement in human disease.Hum Genet. 2021 Mar;140(3):381-400. doi: 10.1007/s00439-020-02212-9. Epub 2020 Jul 29. Hum Genet. 2021. PMID: 32728807 Free PMC article. Review.
-
Pathogenic mechanisms of tooth agenesis linked to paired domain mutations in human PAX9.Hum Mol Genet. 2009 Aug 1;18(15):2863-74. doi: 10.1093/hmg/ddp221. Epub 2009 May 9. Hum Mol Genet. 2009. PMID: 19429910 Free PMC article.
-
PAX8 in the Junction between Development and Tumorigenesis.Int J Mol Sci. 2022 Jul 3;23(13):7410. doi: 10.3390/ijms23137410. Int J Mol Sci. 2022. PMID: 35806410 Free PMC article. Review.
-
Unraveling molecular targets of bisphenol A and S in the thyroid gland.Environ Sci Pollut Res Int. 2018 Sep;25(27):26916-26926. doi: 10.1007/s11356-018-2419-y. Epub 2018 Jul 13. Environ Sci Pollut Res Int. 2018. PMID: 30006815
-
Resonance assignment of disordered protein with repetitive and overlapping sequence using combinatorial approach reveals initial structural propensities and local restrictions in the denatured state.J Biomol NMR. 2016 Sep;66(1):21-35. doi: 10.1007/s10858-016-0054-9. Epub 2016 Sep 1. J Biomol NMR. 2016. PMID: 27586017
References
Publication types
MeSH terms
Substances
Associated data
- Actions
LinkOut - more resources
Full Text Sources
Molecular Biology Databases
Miscellaneous