SRY and human sex determination: The basic tail of the HMG box functions as a kinetic clamp to augment DNA bending

Nelson B. Phillips, Agnes Jancso-Radek, Varda Ittah, Rupinder Singh, Ging Chan, Elisha Haas, Michael A. Weiss

Research output: Contribution to journalArticlepeer-review

33 Scopus citations

Abstract

Human testis-determining factor SRY contains a high-mobility-group (HMG) box, an α-helical DNA-binding domain that binds within an expanded minor groove to induce DNA bending. This motif is flanked on the C-terminal end by a basic tail, which functions both as a nuclear localization signal and accessory DNA-binding element. Whereas the HMG box is broadly conserved among otherwise unrelated transcription factors, tails differ in sequence and mode of DNA binding. Contrasting examples are provided by SRY and lymphoid enhancer factor 1 (LEF-1): whereas the SRY tail remains in the minor groove distal to the HMG box, the LEF-1 tail binds back across the center of the bent DNA site. The LEF-1 tail relieves electrostatic repulsion that would otherwise be incurred within the compressed major groove to enable sharp DNA bending with high affinity. Here, we demonstrate that the analogous SRY tail functions as a "kinetic clamp" to regulate the lifetime of the bent DNA complex. As in LEF-1, partial truncation of the distal SRY tail reduces specific DNA affinity and DNA bending, but these perturbations are modest: binding is reduced by only 1.8-fold, and bending by only 7-10°. "Tailed" and truncated SRY complexes exhibit similar structures (as probed by NMR) and distributions of long-range conformational substates (as probed by time-resolved fluorescence resonance energy transfer). Surprisingly, however, the SRY tail retards dissociation of the protein-DNA complex by 20-fold. The marked and compensating changes in rates of association and dissociation observed on tail truncation, disproportionate to perturbations in affinity or structure, suggest that this accessory element functions as a kinetic clamp to regulate the lifetime of the SRY-DNA complex. We speculate that the kinetic stability of a bent DNA complex is critical to the assembly and maintenance of a sex-specific transcriptional pre-initiation complex.

Original languageEnglish
Pages (from-to)172-192
Number of pages21
JournalJournal of Molecular Biology
Volume358
Issue number1
DOIs
StatePublished - 21 Apr 2006

Bibliographical note

Funding Information:
We thank H. Chen, C.- Y. King; C. Kung, and E. Rivera for participation in early stages of work; N. Narayana for statistical analysis of SRY structures; P. K. Donahoe, Q. X. Hua, D. N. Jones, C.- Y. King, and L. A. Labeots for discussion; and E. Collins and S. Price for preparation of manuscript. This work was supported in part by an equipment grant from the Israel Science Foundation to E.H. (number 553-99), a grant from the United States/Israel Binational Foundation (to E.H. and M.A.W.; number 98–362) and in its early stages by the National Institutes of Health (to M.A.W.; GM051558). These studies are a contribution from the Cleveland Center for Structural Biology.

Funding

We thank H. Chen, C.- Y. King; C. Kung, and E. Rivera for participation in early stages of work; N. Narayana for statistical analysis of SRY structures; P. K. Donahoe, Q. X. Hua, D. N. Jones, C.- Y. King, and L. A. Labeots for discussion; and E. Collins and S. Price for preparation of manuscript. This work was supported in part by an equipment grant from the Israel Science Foundation to E.H. (number 553-99), a grant from the United States/Israel Binational Foundation (to E.H. and M.A.W.; number 98–362) and in its early stages by the National Institutes of Health (to M.A.W.; GM051558). These studies are a contribution from the Cleveland Center for Structural Biology.

FundersFunder number
National Institutes of HealthGM051558
United States-Israel Binational Science Foundation98–362
Israel Science Foundation553-99

    Keywords

    • Gene regulation
    • Gonadal dysgenesis
    • Human development
    • Intersex abnormalities
    • Protein structure

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