Crystallization and X‐ray diffraction analysis of the HMG domain of the chondrogenesis master regulator Sox9 in complex with a ChIP‐Seq‐identified DNA element. Issue 11 (1st November 2015)

• Record Type: Journal Article
• Authors: Vivekanandan, Saravanan; Moovarkumudalvan, Balasubramanian; Lescar, Julien; Kolatkar, Prasanna R.
• Is Part Of: Acta crystallographica. Volume 71:Issue 11(2015:Nov.)
• Page Start: 1437
• Page End: 1441
• Publication Date: 2015-11-01
• Subjects: Sox9 -- transcription factor -- HMG domain -- FOXP2 promoter -- chondrogenesis -- sex‐determining gene; Crystallography -- Periodicals; Crystals -- Periodicals; 548
• DOI: 10.1107/S2053230X1501969X
• Languages: English
• ISSNs: 2053-230X

Abstract

Abstract : Sox9 is a fundamental sex‐determining gene and the master regulator of chondrogenesis, and is involved in the development of various vital organs such as testes, kidney, heart and brain, and in skeletal development. Similar to other known Sox transcription factors, Sox9 recognizes and binds DNA with the consensus sequence C(T/A)TTG(T/A)(T/A) through the highly conserved HMG domain. Nonetheless, the molecular basis of the functional specificity of Sox9 in key developmental processes is still unclear. As an initial step towards a mechanistic understanding of Sox9 transcriptional regulation, the current work describes the details of the purification of the mouse Sox9 HMG domain (mSox9HMG), its crystallization in complex with a ChIP‐Seq‐identified FOXP2 promoter DNA element and the X‐ray diffraction data analysis of this complex. The mSox9HMG–FOXP2 promoter DNA complex was crystallized by the hanging‐drop vapour‐diffusion method using 20% PEG 3350 in 200 m M sodium/potassium phosphate with 100 m M bis‐tris propane at pH 8.5. The crystals diffracted to 2.7 Å resolution and the complex crystallized in the tetragonal space group P 41 21 2, with unit‐cell parameters a = b = 99.49, c = 45.89 Å. Crystal‐packing parameters revealed that asymmetric unit contained one mSox9HMG–FOXP2 promoter DNA complex with an estimated solvent content of 64%.