The LEF-1 high-mobility group domain undergoes a disorder-to-order transition upon formation of a complex with cognate DNA

John J. Love, Xiang Li, John Chung, H. Jane Dyson, Peter E. Wright

Research output: Contribution to journalArticlepeer-review

Abstract

Lymphoid enhancer-binding factor-1 (LEF-1), a member of the high-mobility group (HMG) family of proteins, functions as an architectural transcription factor. In complex with its cognate DNA, the LEF-1 domain is highly ordered, and its NMR spectra are characteristic of a folded globular protein. In contrast, the uncomplexed protein exhibits NMR evidence of substantial conformational heterogeneity, although circular dichroism spectra indicate that much of the α-helical secondary structure of the DNA-bound state is retained in the free protein. Heteronuclear NMR experiments performed on the free LEF-1 domain reveal that helix II and helix III of the HMG domain are folded, although helix III is truncated at its C-terminal end relative to the DNA-bound protein. The major hydrophobic core between helices II and III appears to be formed, but the minor core near the C-terminus of helix III is unstructured in the free protein. Backbone resonances of helix I are undetectable, probably as a result of exchange broadening due to fluctuations between two or more conformations on a microsecond-to-millisecond time scale. On the basis of the circular dichroism spectrum, this region of the polypeptide appears to adopt helical structure but the helix is not fully stabilized in the absence of DNA. These findings argue that, prior to binding, bending, and distorting DNA, the HMG domain of LEF-1 exists in a segmentally disordered or partially folded state. Upon complex formation, the protein domain undergoes a cooperative folding transition with DNA to a highly ordered and well-folded state.

Original languageEnglish (US)
Pages (from-to)8725-8734
Number of pages10
JournalBiochemistry
Volume43
Issue number27
DOIs
StatePublished - Jul 13 2004

ASJC Scopus subject areas

  • Biochemistry

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