Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1992 Nov 1;89(21):10479–10483. doi: 10.1073/pnas.89.21.10479

Supercoiled DNA wraps around the bacteriophage phi 29 head-tail connector.

S Turnquist 1, M Simon 1, E Egelman 1, D Anderson 1
PMCID: PMC50362  PMID: 1438237

Abstract

Supercoiled pBR322 DNA wraps around the outside of the isolated Bacillus subtilis bacteriophage phi 29 head-tail connector, the crux of the DNA packaging machine of the viral precursor capsid or prohead. The contour length of the supercoiled DNA, determined by EM, decreased by approximately 180 base pairs for each connector bound. Mass and radial density determinations by scanning transmission EM showed that the increased mass of the connector-DNA complex relative to the connector alone was equivalent to approximately 170 base pairs of DNA and was located around the outside of the connector. Topoisomerase I treatment of the complexes followed by deproteinization suggested that supercoils were restrained by the connectors. Connectors bound linear and open-circular plasmid DNAs inefficiently but were not wrapped by these DNAs. The wrapping of supercoiled DNA around the isolated phi 29 connector is hypothesized to reflect the initiation phase of the normal process of DNA packaging. Packaging substrates would be supercoiled, wrapped by the connector, linearized, and translocated by rotation of the connector relative to the viral capsid with the aid of ATP hydrolysis.

Full text

PDF
10479

Images in this article

10480Image
on p.10480
10480Image
on p.10480
10481Image
on p.10481
10482Image
on p.10482

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bazinet C., King J. The DNA translocating vertex of dsDNA bacteriophage. Annu Rev Microbiol. 1985;39:109–129. doi: 10.1146/annurev.mi.39.100185.000545. [DOI] [PubMed] [Google Scholar]
  2. Better M., Lu C., Williams R. C., Echols H. Site-specific DNA condensation and pairing mediated by the int protein of bacteriophage lambda. Proc Natl Acad Sci U S A. 1982 Oct;79(19):5837–5841. doi: 10.1073/pnas.79.19.5837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bjornsti M. A., Reilly B. E., Anderson D. L. Morphogenesis of bacteriophage phi 29 of Bacillus subtilis: oriented and quantized in vitro packaging of DNA protein gp3. J Virol. 1983 Jan;45(1):383–396. doi: 10.1128/jvi.45.1.383-396.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Carazo J. M., Donate L. E., Herranz L., Secilla J. P., Carrascosa J. L. Three-dimensional reconstruction of the connector of bacteriophage phi 29 at 1.8 nm resolution. J Mol Biol. 1986 Dec 20;192(4):853–867. doi: 10.1016/0022-2836(86)90033-1. [DOI] [PubMed] [Google Scholar]
  5. Donate L. E., Herranz L., Secilla J. P., Carazo J. M., Fujisawa H., Carrascosa J. L. Bacteriophage T3 connector: three-dimensional structure and comparison with other viral head-tail connecting regions. J Mol Biol. 1988 May 5;201(1):91–100. doi: 10.1016/0022-2836(88)90441-x. [DOI] [PubMed] [Google Scholar]
  6. Fuller R. S., Funnell B. E., Kornberg A. The dnaA protein complex with the E. coli chromosomal replication origin (oriC) and other DNA sites. Cell. 1984 Oct;38(3):889–900. doi: 10.1016/0092-8674(84)90284-8. [DOI] [PubMed] [Google Scholar]
  7. García J. A., Méndez E., Salas M. Cloning, nucleotide sequence and high level expression of the gene coding for the connector protein of Bacillus subtilis phage phi 29. Gene. 1984 Oct;30(1-3):87–98. doi: 10.1016/0378-1119(84)90108-2. [DOI] [PubMed] [Google Scholar]
  8. Grimes S., Anderson D. RNA dependence of the bacteriophage phi 29 DNA packaging ATPase. J Mol Biol. 1990 Oct 20;215(4):559–566. doi: 10.1016/s0022-2836(05)80168-8. [DOI] [PubMed] [Google Scholar]
  9. Guo P. X., Erickson S., Anderson D. A small viral RNA is required for in vitro packaging of bacteriophage phi 29 DNA. Science. 1987 May 8;236(4802):690–694. doi: 10.1126/science.3107124. [DOI] [PubMed] [Google Scholar]
  10. Guo P., Grimes S., Anderson D. A defined system for in vitro packaging of DNA-gp3 of the Bacillus subtilis bacteriophage phi 29. Proc Natl Acad Sci U S A. 1986 May;83(10):3505–3509. doi: 10.1073/pnas.83.10.3505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hendrix R. W. Symmetry mismatch and DNA packaging in large bacteriophages. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4779–4783. doi: 10.1073/pnas.75.10.4779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ibáez C., García J. A., Carrascosa J. L., Salas M. Overproduction and purification of the connector protein of Bacillus subtilis phage phi 29. Nucleic Acids Res. 1984 Mar 12;12(5):2351–2365. doi: 10.1093/nar/12.5.2351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kirchhausen T., Wang J. C., Harrison S. C. DNA gyrase and its complexes with DNA: direct observation by electron microscopy. Cell. 1985 Jul;41(3):933–943. doi: 10.1016/s0092-8674(85)80074-x. [DOI] [PubMed] [Google Scholar]
  14. Richmond T. J., Finch J. T., Rushton B., Rhodes D., Klug A. Structure of the nucleosome core particle at 7 A resolution. Nature. 1984 Oct 11;311(5986):532–537. doi: 10.1038/311532a0. [DOI] [PubMed] [Google Scholar]
  15. Zahn K., Blattner F. R. Direct evidence for DNA bending at the lambda replication origin. Science. 1987 Apr 24;236(4800):416–422. doi: 10.1126/science.2951850. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES