Characterization of DNA–Hv1 histone interactions; discrimination of DNA size and shape

Edited by Richard Cogdell

Received 29 September 2009; received in revised form 14 January 2010; accepted 14 January 2010. published online 18 January 2010.

Abstract

We have studied the formation of histone Hv1–DNA complexes using an acoustic biosensor and AFM imaging. Our results show that DNA and histone molecules aggregate into amorphous accumulations which form a compact rigid layer on the sensor’s surface. By measuring changes in the acoustic wave amplitude, it was possible to titrate surface bound DNA with Hv1 and discriminate between DNA molecules of different size and shape. From the kinetic analysis of real time data, Keq was found equal to 3×105M−1.

Keywords: Acoustic biosensor, Hv1 histone protein, DNA shape, Nucleoprotein complex, Rigid layer

a Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology Hellas, Vassilika Vouton, 71110 Heraklion, Greece

b Department of Materials Science and Technology, University of Crete, Vassilika Vouton, 71003 Heraklion, Greece

c Department of Biology, University of Crete, Vassilika Vouton, 71409 Heraklion, Greece

Corresponding author. Address: University of Crete, Department of Biology, Vassilika Vouton, 394408 Heraklion, Greece. Fax: +30 2810 394408.

PII: S0014-5793(10)00053-0

doi:10.1016/j.febslet.2010.01.030

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