The Myb domain of LUX ARRHYTHMO in complex with DNA: expression, purification and crystallization

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• 作者：Catarina S. Silva ; Xuelei Lai ; Max Nanao and Chloe Zubieta
• 关键词：Myb domain ; DNA binding ; protein&ndash ; DNA complex ; LUX ARRHYTHMO ; Arabidopsis thaliana
• 刊名：Acta Crystallographica Section F
• 出版年：2016
• 出版时间：May 2016
• 年：2016
• 卷：72
• 期：5
• 页码：356-361
• 全文大小：1061K
• ISSN：1744-3091

文摘

LUX ARRHYTHMO (LUX) is a Myb-domain transcription factor that plays an important role in regulating the circadian clock. Lux mutations cause severe clock defects and arrhythmia in constant light and dark. In order to examine the molecular mechanisms underlying the function of LUX, the DNA-binding Myb domain was cloned, expressed and purified. The DNA-binding activity of the Myb domain was confirmed using electrophoretic mobility shift assays (EMSAs), demonstrating that the LUX Myb domain is able to bind to DNA with nanomolar affinity. In order to investigate the specificity determinants of protein–DNA interactions, the protein was co-crystallized with a 10-mer cognate DNA. Initial crystallization results for the selenomethionine-derivatized protein and data-set collection statistics are reported. Data collection was performed using the MeshAndCollect workflow available at the ESRF.