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Related Experiment Videos

Bent helical structure in kinetoplast DNA.

J C Marini1, S D Levene, D M Crothers

  • 1Department of Physiological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.

Proceedings of the National Academy of Sciences of the United States of America
|December 1, 1982
PubMed
Summary

Leishmania tarentolae kinetoplast DNA exhibits unusual physical properties due to its unique sequence. This DNA fragment displays anomalous behavior in various gel electrophoresis and filtration experiments, suggesting a compact, systematically bent B-DNA structure.

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Area of Science:

  • Molecular Biology
  • Biophysics
  • Genetics

Background:

  • Kinetoplast DNA (kDNA) from Leishmania tarentolae is a unique genetic material.
  • Understanding DNA structure-function relationships is crucial in molecular biology.

Purpose of the Study:

  • To investigate the anomalous physical properties of a specific Leishmania tarentolae kDNA restriction fragment.
  • To elucidate the structural basis for the observed anomalous behavior.

Main Methods:

  • Maxam-Gilbert sequencing to determine fragment length.
  • Agarose and polyacrylamide gel electrophoresis to assess electrophoretic behavior.
  • Sephacryl S-500 gel filtration to evaluate molecular size.
  • Transient electric dichroism studies to measure rotational relaxation times.

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Main Results:

  • A 490 bp kDNA fragment showed anomalous migration patterns in gel electrophoresis (450 bp vs. 1,380 bp) and gel filtration (375 bp).
  • Electric dichroism experiments revealed a smaller rotational relaxation time compared to a control fragment.
  • Control experiments ruled out DNA modification as the cause of these properties.

Conclusions:

  • The anomalous behavior is attributed to the DNA sequence itself, leading to a compact configuration.
  • A model of systematically bent B-DNA is proposed to explain the observed properties.
  • Periodicities within the DNA sequence are suggested as the cause of DNA bending.