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Initiation events in in-vitro packaging of bacteriophage phi 29 DNA-gp3.

Abstract

Initiation events in the packaging of bacteriophage phi 29 DNA-gp3 (DNA-gene product 3 complex) were studied in a completely defined in-vitro system that included purified proheads, DNA-gp3 and the DNA packaging protein gp16. In the sequential interactions, gp16 first bound to, and was modified by, the prohead. The prohead-gp16 complex then bound to DNA-gp3, resulting in a second modification of gp16 that permitted binding of ATP. DNA-gp3 aggregates were produced, and the hydrolysis of ATP accompanied DNA-gp3 packaging. Binding and hydrolysis of ATP by gp16 was both prohead- and DNA-gp3-dependent. Interruption of packaging by DNase I addition revealed filled heads but few particles containing partial lengths of DNA, suggesting that following a rate-limiting initiation, the translocation of DNA-gp3 into the prohead was much faster in the defined in-vitro system than in extracts.

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  • Publisher Full Text
  • Authors+Show Affiliations

    ,

    Department of Microbiology, University of Minnesota, Minneapolis 55455.

    ,

    Source

    Journal of molecular biology 197:2 1987 Sep 20 pg 219-28

    MeSH

    Adenosine Triphosphate
    Bacillus subtilis
    Bacteriophages
    Centrifugation, Density Gradient
    DNA, Viral
    Microscopy, Electron
    Viral Proteins

    Pub Type(s)

    Journal Article
    Research Support, U.S. Gov't, P.H.S.

    Language

    eng

    PubMed ID

    3119862

    Citation

    Guo, P, et al. "Initiation Events in In-vitro Packaging of Bacteriophage Phi 29 DNA-gp3." Journal of Molecular Biology, vol. 197, no. 2, 1987, pp. 219-28.
    Guo P, Peterson C, Anderson D. Initiation events in in-vitro packaging of bacteriophage phi 29 DNA-gp3. J Mol Biol. 1987;197(2):219-28.
    Guo, P., Peterson, C., & Anderson, D. (1987). Initiation events in in-vitro packaging of bacteriophage phi 29 DNA-gp3. Journal of Molecular Biology, 197(2), pp. 219-28.
    Guo P, Peterson C, Anderson D. Initiation Events in In-vitro Packaging of Bacteriophage Phi 29 DNA-gp3. J Mol Biol. 1987 Sep 20;197(2):219-28. PubMed PMID: 3119862.
    * Article titles in AMA citation format should be in sentence-case
    TY - JOUR T1 - Initiation events in in-vitro packaging of bacteriophage phi 29 DNA-gp3. AU - Guo,P, AU - Peterson,C, AU - Anderson,D, PY - 1987/9/20/pubmed PY - 1987/9/20/medline PY - 1987/9/20/entrez SP - 219 EP - 28 JF - Journal of molecular biology JO - J. Mol. Biol. VL - 197 IS - 2 N2 - Initiation events in the packaging of bacteriophage phi 29 DNA-gp3 (DNA-gene product 3 complex) were studied in a completely defined in-vitro system that included purified proheads, DNA-gp3 and the DNA packaging protein gp16. In the sequential interactions, gp16 first bound to, and was modified by, the prohead. The prohead-gp16 complex then bound to DNA-gp3, resulting in a second modification of gp16 that permitted binding of ATP. DNA-gp3 aggregates were produced, and the hydrolysis of ATP accompanied DNA-gp3 packaging. Binding and hydrolysis of ATP by gp16 was both prohead- and DNA-gp3-dependent. Interruption of packaging by DNase I addition revealed filled heads but few particles containing partial lengths of DNA, suggesting that following a rate-limiting initiation, the translocation of DNA-gp3 into the prohead was much faster in the defined in-vitro system than in extracts. SN - 0022-2836 UR - https://www.unboundmedicine.com/medline/citation/3119862/Initiation_events_in_in_vitro_packaging_of_bacteriophage_phi_29_DNA_gp3_ L2 - https://linkinghub.elsevier.com/retrieve/pii/0022-2836(87)90120-3 DB - PRIME DP - Unbound Medicine ER -