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Optical tweezers reveal force plateau and internal friction in PEG-induced DNA condensation
Authors:Heikki Ojala  Gabija Ziedaite  Anders E. Wallin  Dennis H. Bamford  Edward Hæggström
Affiliation:1. Department of Physics, University of Helsinki, Gustaf H?llstr?min katu 2, P.O. Box 64, 00014, Helsinki, Finland
2. Department of Biosciences, University of Helsinki, Viikinkaari 5, P.O. Box 56, 00014, Helsinki, Finland
3. Centre for Metrology and Accreditation, Tekniikantie 1, P.O. Box 9, 02151, Espoo, Finland
4. Institute of Biotechnology, University of Helsinki, Viikinkaari 5, P.O. Box 56, 00014, Helsinki, Finland
Abstract:
The simplified artificial environments in which highly complex biological systems are studied do not represent the crowded, dense, salty, and dynamic environment inside the living cell. Consequently, it is important to investigate the effect of crowding agents on DNA. We used a dual-trap optical tweezers instrument to perform force spectroscopy experiments at pull speeds ranging from 0.3 to 270 μm/s on single dsDNA molecules in the presence of poly(ethylene glycol) (PEG) and monovalent salt. PEG of sizes 1,500 and 4,000 Da condensed DNA, and force–extension data contained a force plateau at approximately 1 pN. The level of the force plateau increased with increasing pull speed. During slow pulling the dissipated work increased linearly with pull speed. The calculated friction coefficient did not depend on amount of DNA incorporated in the condensate, indicating internal friction is independent of the condensate size. PEG300 had no effect on the dsDNA force–extension curve. The force plateau implies that condensation induced by crowding agents resembles condensation induced by multivalent cations.
Keywords:
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