Identification of a Pax paired domain recognition sequence and evidence for DNA-dependent conformational changes

Jonathan Epstein, Jiexing Cai, Thomas M Glaser, Lisa Jepeal, Richard Maas

Research output: Contribution to journalArticle

295 Citations (Scopus)

Abstract

Pax genes encode a family of developmentally regulated transcription factors that have been implicated in a number of human and murine congenital disorders, as well as in tumorigenesis (Gruss, P., and Walther, C. (1992) Cell 69, 719-722; Hill, R., and van Heyningen, V. (1992) Trends Genet. 8, 119-120; Chalepakis, G., Tremblay, P., and Gruss, P. (1992) J. Cell Sci. Suppl. 16, 61-67; Maulbecker, C. C., and Gruss, P. (1993) EMBO J. 12, 2361- 2367; Walther, C., Guenet, J. L., Simon, D., Deutsch, U., Jostes, B., Goulding, M. D., Plachov, D., Balling, R., and Gruss, P. (1991) Genomics 11, 424-434; Barr, R. G., Galili, N., Holick, J., Biegel, J. A., Rovera, G., and Emanuel, B. S. (1993) Nature Genet. 3, 113-117). These genes are defined by the presence of an evolutionarily conserved DNA binding domain, termed the paired domain. The structure and the DNA binding characteristics of the paired domain remain largely unknown. We have utilized repetitive rounds of a polymerase chain reaction-based selection method to identify the optimal DNA binding sequences for the Pax-2 and Pax-6 paired domains. The results suggest that the paired domain family of peptides bind similar DNA sequences. Identification of this binding site has revealed an important structural clue regarding the mechanism of paired domain binding to DNA. CD and NMR structural analyses of the purified Pax-6 paired domain reveal it to be largely structureless in solution. Upon binding the recognition sequence, the complex becomes markedly less soluble and displays CD spectroscopic evidence of significant α-helical structure.

Original languageEnglish (US)
Pages (from-to)8355-8361
Number of pages7
JournalJournal of Biological Chemistry
Volume269
Issue number11
StatePublished - Mar 18 1994
Externally publishedYes

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Viverridae
DNA
Congenital, Hereditary, and Neonatal Diseases and Abnormalities
Genes
Genomics
Polymerase chain reaction
DNA sequences
Carcinogenesis
Transcription Factors
Binding Sites
Polymerase Chain Reaction
Nuclear magnetic resonance
Peptides
Protein Domains

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification of a Pax paired domain recognition sequence and evidence for DNA-dependent conformational changes. / Epstein, Jonathan; Cai, Jiexing; Glaser, Thomas M; Jepeal, Lisa; Maas, Richard.

In: Journal of Biological Chemistry, Vol. 269, No. 11, 18.03.1994, p. 8355-8361.

Research output: Contribution to journalArticle

Epstein, Jonathan ; Cai, Jiexing ; Glaser, Thomas M ; Jepeal, Lisa ; Maas, Richard. / Identification of a Pax paired domain recognition sequence and evidence for DNA-dependent conformational changes. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 11. pp. 8355-8361.
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abstract = "Pax genes encode a family of developmentally regulated transcription factors that have been implicated in a number of human and murine congenital disorders, as well as in tumorigenesis (Gruss, P., and Walther, C. (1992) Cell 69, 719-722; Hill, R., and van Heyningen, V. (1992) Trends Genet. 8, 119-120; Chalepakis, G., Tremblay, P., and Gruss, P. (1992) J. Cell Sci. Suppl. 16, 61-67; Maulbecker, C. C., and Gruss, P. (1993) EMBO J. 12, 2361- 2367; Walther, C., Guenet, J. L., Simon, D., Deutsch, U., Jostes, B., Goulding, M. D., Plachov, D., Balling, R., and Gruss, P. (1991) Genomics 11, 424-434; Barr, R. G., Galili, N., Holick, J., Biegel, J. A., Rovera, G., and Emanuel, B. S. (1993) Nature Genet. 3, 113-117). These genes are defined by the presence of an evolutionarily conserved DNA binding domain, termed the paired domain. The structure and the DNA binding characteristics of the paired domain remain largely unknown. We have utilized repetitive rounds of a polymerase chain reaction-based selection method to identify the optimal DNA binding sequences for the Pax-2 and Pax-6 paired domains. The results suggest that the paired domain family of peptides bind similar DNA sequences. Identification of this binding site has revealed an important structural clue regarding the mechanism of paired domain binding to DNA. CD and NMR structural analyses of the purified Pax-6 paired domain reveal it to be largely structureless in solution. Upon binding the recognition sequence, the complex becomes markedly less soluble and displays CD spectroscopic evidence of significant α-helical structure.",
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