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Electron microscopic mapping of proteins bound to herpes simplex virus DNA

Wu, Madeline and Hyman, Richard W. and Davidson, Norman (1979) Electron microscopic mapping of proteins bound to herpes simplex virus DNA. Nucleic Acids Research, 6 (11). pp. 3427-3441. ISSN 0305-1048. http://resolver.caltech.edu/CaltechAUTHORS:WUMnar79

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Abstract

Exonuclease digestion experiments have suggested that there is a protein(s) bound close to one or both ends of herpes sir virus-1 (HSV) DNA. The existence of such bound proteins has been positiviely demonstrated and their positions on the HSV genolie determined by application of a newly developed method for electron microscopic mapping of proteins bound to nucleic acids. Purified HSV DNA was treated with dinitrofluorobenzene under conditions that covalently attach the dinitrophenyl (DNP) group to the proteins in protein-nucleic acid complex. The HSV DNA-protein-(DNP)n complex was treated with rabbit anti-DNP IgG, and, in some cases, additionally treated with monovalent Fab fragments of goat anti-rabbit IgG, and mounted for examination in the electron microscope. Electron opaque dots representing the protein-(DNP)n-(IgG)m complex were seen on the HSV DNA. Direct measurements of the positions of the protein, as well as partial denaturation mapping, indicate that there are four positions for protein bound to HSV DNA: two near but not at the two ends and two at sites corresponding to the internalinverted repeats of the ends. These results suggest that there is a specific protein binding sequence within the direct terminal repeat of HSV DNA. The previous obseroation t HSV DNA is more sensitive to digestion by a 3' than by a 5' exonuolease then indicates that the bound protein(s) is more intimately associated with one strand of the specific sequence than with the complementary strand.


Item Type:Article
Additional Information:Copyright © 1979 Oxford University Press. Received June 4, 1979. M.W. and N.D. acknowledge support by Grant GM 20927 with the United States Public Health Service. We thank Mrs. Linda Kudler for her excellent technical assistance. R.W.H. is supported by Contract #NO1 CP 5 3516 with the Virus Cancer Program of the National Cancer Institute, by Grants CA 16498 and CA 18450 awarded by the NCI, and is the recipient of a Faculty Research Award from the American Cancer Society (FRA-158).
Record Number:CaltechAUTHORS:WUMnar79
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:WUMnar79
Alternative URL:http://nar.oxfordjournals.org/cgi/content/abstract/6/11/3427
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:4059
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:26 Jul 2006
Last Modified:26 Dec 2012 08:57

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