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J Virol. 1977 August; 23(2): 368-376
Copyright © 1977 American Society for Microbiology. All Rights Reserved.

Structure of Hepatitis B Dane Particle DNA and Nature of the Endogenous DNA Polymerase Reaction

Terry A. Landers1, Harry B. Greenberg2 and William S. Robinson

a Departments of Medicine and Biochemistry, Stanford University School of Medicine, Stanford, California 94305

ABSTRACT

The circular DNA of hepatitis B Dane particles, which serves as the primer/template for an endogenous DNA polymerase, was analyzed by electrophoresis before and after a polymerase reaction and after digestion by restriction endonuclease or single-strand-specific endonuclease S1. The unreacted molecules extracted from the particles were electrophoretically heterogeneous, and treatment with S1 nuclease produced double-stranded linear DNA ranging in length from 1,700 to 2,800 base pairs (bp). After an endogenous DNA polymerase reaction, two discrete species of DNA molecules were found: a circular form and a linear form 3,200 bp long. The reaction resulted in a population of molecules with an elongated and more homogeneous double-stranded region. These results suggest that the circular molecules in Dane particles have single-stranded regions of varying lengths that are made double stranded during the DNA polymerase reaction. The endogenous DNA polymerase was found to initiate apparently at random in a region spanning more than a third of the molecule. Analysis of restriction endonuclease cleavage fragments of the fully elongated DNA revealed that although the molecules were of a uniform length, they were somewhat heterogeneous in sequence. The sum of the sizes of the 10 major endonuclease Hae III-generated fragments, detected by ethidium bromide, was 3,880 bp. Two additional fragments (B and G) detected by autoradiography after an endogenous DNA polymerase reaction with 32P-labeled deoxynucleoside triphosphates made the total 4,910 bp.

FOOTNOTES

1 Present address: Department of Biochemistry and Molecular Biology, University of Texas Medical School, Houston, TX 77030.

2 Present address: Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20014.

J Virol. 1977 August; 23(2): 368-376
Copyright © 1977 American Society for Microbiology. All Rights Reserved.



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