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Journal of Bacteriology, February 2000, p. 680-688, Vol. 182, No. 3
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Bacteriophage T4 Self-Assembly: Localization of gp3 and Its Role in Determining Tail Length

A. Vianelli,^1, G. R. Wang,^1, M. Gingery,² R. L. Duda,^2,§ F. A. Eiserling,^2,* and E. B. Goldberg^1,*

Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts 02111,¹ and Department of Microbiology and Molecular Genetics, University of California at Los Angeles, Los Angeles, California 90024²

Received 24 June 1996/Accepted 15 October 1999

Gene 3 of bacteriophage T4 participates at a late stage in the T4 tail assembly pathway, but the hypothetical protein product, gp3, has never been identified in extracts of infected cells or in any tail assembly intermediate. In order to overcome this difficulty, we expressed gp3 in a high-efficiency plasmid expression vector and subsequently purified it for further analysis. The N-terminal sequence of the purified protein showed that the initial methionine had been removed. Variant C-terminal amino acid sequences were resolved by determining the cysteine content of the protein. The molecular mass of 20.6 kDa for the pure protein was confirmed by Western blotting, using a specific anti-gp3 serum for which the purified protein was the immunogen. We also demonstrated, for the first time, the physical presence of gp3 in the mature T4 phage particle and localized it to the tail tube. By finding a nonleaky, nonpermissive host for a gene 3 mutant, we could clearly demonstrate a new phenotype: the slow, aberrant elongation of the tail tube in the absence of gp3.

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^* Corresponding author. Mailing address for E. B. Goldberg: Department of Molecular Biology and Microbiology, Tufts University School of Medicine, 136 Harrison Ave., Boston, MA 02111. Phone: (617) 636-6754. Fax: (617) 636-0337. E-mail: egoldber{at}opal.tufts.edu. Mailing address for F. A. Eiserling: Department of Microbiology and Molecular Genetics, University of California at Los Angeles, Los Angeles, CA 90024. Phone: (310) 825-4959. Fax: (310) 825-9368. E-mail: frede{at}college.UCLA.edu.

Present address: Department of Structural and Functional Biology, University of Insubria, Via J. H. Durant 3, 21100 Varese, Italy.

Present address: Molecular Cardiology Research Institute, New England Medical Center, Boston, MA 02111.

^§ Present address: Department of Biological Science, University of Pittsburgh, Pittsburgh, PA 15260.

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Journal of Bacteriology, February 2000, p. 680-688, Vol. 182, No. 3
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

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