A quantitative test of Zimm's model for the rotor‐speed‐department sedimentation of linear DNA molecules
- 1 May 1980
- journal article
- research article
- Published by Wiley in Biopolymers
- Vol. 19 (5) , 945-964
- https://doi.org/10.1002/bip.1980.360190502
Abstract
In 1974, Zimm described a theory which predicts that the sedimentation coefficient of high‐molecular‐weight DNA will decrease as the rotor speed of measurement increases. In 1979, this theory was revised, and the new formula predicts speed‐dependence effects that are substantially smaller than the predictions of the original version. This report describes the results of subjecting both the original and the revised versions of the theory to quantitative tests using a well‐defined sucrose‐gradient system and a DNA of known molecular weight (T4c DNA). T4c bacteriophage is a mutant, whose DNA contains the unmodified base cytosine, instead of the glucosylated hydroxymethylcytosine characteristic of the T‐even bacteriophages, and has a molecular weight of 115 ± 3 × 106. The DNA of the wild‐type phage (T4D+) was also used in some experiments.In addition to the quantitative tests, the experiments test for an effect first observed by Rubenstein and Leighton, which showed that the sedimentation coefficient measured for T2 DNA depended on the composition of the centrifuge tube used for the measurement (tube composition effect). It can be inferred from this observation that an interaction occurs between particle and tube wall during sedimentation, and this leads to a reduction in sedimentation velocity independent of the reduction in S described by Zimm's theory.The results show that in the range of 25,000–50,000 rpm, the original but theoretically incorrect form of the theory quite accurately describes the sedimentation behavior of both T4c and T4D+ DNA, although T4D+ was a special case in some respects. The revised (corrected) form of the theory predicts much less of a speed‐dependence effect than that actually observed. The discrepancy between corrected theory and observation suggests that other factors (perhaps arising from the use of the swinging bucket rotor geometry) are causing the additional observed reduction in S20,w. However, the experiments show that the tube composition effect does not seem to be one of these.This publication has 24 references indexed in Scilit:
- Density‐gradient‐sedimentation velocity of solvated macromolecules: Theoretical considerations about the buoyancy factorBiopolymers, 1979
- An explanation for rotor speed effects observed during sedimentation of large folded DNA moleculesJournal of Molecular Biology, 1978
- Repair of DNA double-strand breaks in Escherichia coli, which requires recA function and the presence of a duplicate genomeJournal of Molecular Biology, 1977
- Sedimentation properties of the bacterial chromosome as an isolated nucleoid and as an unfolded DNA fiberJournal of Molecular Biology, 1977
- Bovine serum albumin in aqueous guanidine hydrochloride solutions. Preferential and absolute interactions and comparison with other systemsBiochemistry, 1977
- The production of undegraded cytosine-containing DNA by bacteriophage T4 in the absence of dCTPase and endonucleases II and IV, and its effects on T4-directed protein synthesisJournal of Molecular Biology, 1975
- A rotor speed dependent crossover in sedimentation velocities of DNA's of different sizesBiochemical and Biophysical Research Communications, 1974
- Neutral sucrose sedimentation of very large DNA from Bacillus subtilis: I. Effect of random double-strand breaks and centrifuge speed on sedimentationJournal of Molecular Biology, 1973
- Characterization of the Bacillus subtilis W23 genome by sedimentationJournal of Molecular Biology, 1972
- Sedimentation Coefficients of Linear and Cyclic Wormlike Coils with Excluded-Volume EffectsThe Journal of Chemical Physics, 1967