Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1993 Oct;67(10):6327–6331. doi: 10.1128/jvi.67.10.6327-6331.1993

Characterization of the DNA binding properties of polyomavirus capsid protein.

D Chang 1, X Cai 1, R A Consigli 1
PMCID: PMC238062  PMID: 8396688

Abstract

The DNA binding properties of the polyomavirus structural proteins VP1, VP2, and VP3 were studied by Southwestern analysis. The major viral structural protein VP1 and host-contributed histone proteins of polyomavirus virions were shown to exhibit DNA binding activity, but the minor capsid proteins VP2 and VP3 failed to bind DNA. The N-terminal first five amino acids (Ala-1 to Lys-5) were identified as the VP1 DNA binding domain by genetic and biochemical approaches. Wild-type VP1 expressed in Escherichia coli (RK1448) exhibited DNA binding activity, but the N-terminal truncated VP1 mutants (lacking Ala-1 to Lys-5 and Ala-1 to Cys-11) failed to bind DNA. The synthetic peptide (Ala-1 to Cys-11) was also shown to have an affinity for DNA binding. Site-directed mutagenesis of the VP1 gene showed that the point mutations at Pro-2, Lys-3, and Arg-4 on the VP1 molecule did not affect DNA binding properties but that the point mutation at Lys-5 drastically reduced DNA binding affinity. The N-terminal (Ala-1 to Lys-5) region of VP1 was found to be essential and specific for DNA binding, while the DNA appears to be non-sequence specific. The DNA binding domain and the nuclear localization signal are located in the same N-terminal region.

Full text

PDF
6327

Images in this article

6328Image
on p.6328
6329Image
on p.6329
6329Image
on p.6329
6329Image
on p.6329
6330Image
on p.6330

Selected References

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

  1. Bolen J. B., Anders D. G., Trempy J., Consigli R. A. Differences in the subpopulations of the structural proteins of polyoma virions and capsids: biological functions of the multiple VP1 species. J Virol. 1981 Jan;37(1):80–91. doi: 10.1128/jvi.37.1.80-91.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bowen B., Steinberg J., Laemmli U. K., Weintraub H. The detection of DNA-binding proteins by protein blotting. Nucleic Acids Res. 1980 Jan 11;8(1):1–20. doi: 10.1093/nar/8.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brady J. N., Winston V. D., Consigli R. A. Characterization of a DNA-protein complex and capsomere subunits derived from polyoma virus by treatment with ethyleneglycol-bis-N,N'-tetraacetic acid and dithiothreitol. J Virol. 1978 Jul;27(1):193–204. doi: 10.1128/jvi.27.1.193-204.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chang D., Haynes J. I., 2nd, Brady J. N., Consigli R. A. Identification of a nuclear localization sequence in the polyomavirus capsid protein VP2. Virology. 1992 Dec;191(2):978–983. doi: 10.1016/0042-6822(92)90276-u. [DOI] [PubMed] [Google Scholar]
  5. Chang D., Haynes J. I., 2nd, Brady J. N., Consigli R. A. The use of additive and subtractive approaches to examine the nuclear localization sequence of the polyomavirus major capsid protein VP1. Virology. 1992 Aug;189(2):821–827. doi: 10.1016/0042-6822(92)90615-v. [DOI] [PubMed] [Google Scholar]
  6. Chang D., Paulsen A., Johnson T. C., Consigli R. A. Virus protein assembly in microgravity. Adv Space Res. 1993;13(7):251–257. doi: 10.1016/0273-1177(93)90380-t. [DOI] [PubMed] [Google Scholar]
  7. Delos S. E., Montross L., Moreland R. B., Garcea R. L. Expression of the polyomavirus VP2 and VP3 proteins in insect cells: coexpression with the major capsid protein VP1 alters VP2/VP3 subcellular localization. Virology. 1993 May;194(1):393–398. doi: 10.1006/viro.1993.1274. [DOI] [PubMed] [Google Scholar]
  8. Fattaey A. R., Consigli R. A. Synthesis, posttranslational modifications, and nuclear transport of polyomavirus major capsid protein VP1. J Virol. 1989 Jul;63(7):3168–3175. doi: 10.1128/jvi.63.7.3168-3175.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Forstová J., Krauzewicz N., Wallace S., Street A. J., Dilworth S. M., Beard S., Griffin B. E. Cooperation of structural proteins during late events in the life cycle of polyomavirus. J Virol. 1993 Mar;67(3):1405–1413. doi: 10.1128/jvi.67.3.1405-1413.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Garcea R. L., Benjamin T. L. Isolation and characterization of polyoma nucleoprotein complexes. Virology. 1983 Oct 15;130(1):65–75. doi: 10.1016/0042-6822(83)90118-6. [DOI] [PubMed] [Google Scholar]
  11. Griffith J. P., Griffith D. L., Rayment I., Murakami W. T., Caspar D. L. Inside polyomavirus at 25-A resolution. Nature. 1992 Feb 13;355(6361):652–654. doi: 10.1038/355652a0. [DOI] [PubMed] [Google Scholar]
  12. Haynes J. I., 2nd, Chang D., Consigli R. A. Mutations in the putative calcium-binding domain of polyomavirus VP1 affect capsid assembly. J Virol. 1993 May;67(5):2486–2495. doi: 10.1128/jvi.67.5.2486-2495.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Liddington R. C., Yan Y., Moulai J., Sahli R., Benjamin T. L., Harrison S. C. Structure of simian virus 40 at 3.8-A resolution. Nature. 1991 Nov 28;354(6351):278–284. doi: 10.1038/354278a0. [DOI] [PubMed] [Google Scholar]
  14. McMillen J., Consigli R. A. Characterization of polyoma DNA-protein complexes. I. Electrophoretic identification of the proteins in a nucleoprotein complex isolated from polyoma-infected cells. J Virol. 1974 Dec;14(6):1326–1336. doi: 10.1128/jvi.14.6.1326-1336.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. McMillen J., Consigli R. A. Immunological reactivity of antisera to sodium dodecyl sulfate-derived polypeptides of polyoma virions. J Virol. 1977 Mar;21(3):1113–1120. doi: 10.1128/jvi.21.3.1113-1120.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Merrifield B. Solid phase synthesis. Science. 1986 Apr 18;232(4748):341–347. doi: 10.1126/science.3961484. [DOI] [PubMed] [Google Scholar]
  17. Moreland R. B., Garcea R. L. Characterization of a nuclear localization sequence in the polyomavirus capsid protein VP1. Virology. 1991 Nov;185(1):513–518. doi: 10.1016/0042-6822(91)90811-o. [DOI] [PubMed] [Google Scholar]
  18. Moreland R. B., Montross L., Garcea R. L. Characterization of the DNA-binding properties of the polyomavirus capsid protein VP1. J Virol. 1991 Mar;65(3):1168–1176. doi: 10.1128/jvi.65.3.1168-1176.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Salunke D. M., Caspar D. L., Garcea R. L. Self-assembly of purified polyomavirus capsid protein VP1. Cell. 1986 Sep 12;46(6):895–904. doi: 10.1016/0092-8674(86)90071-1. [DOI] [PubMed] [Google Scholar]
  20. Soussi T. DNA-binding properties of the major structural protein of simian virus 40. J Virol. 1986 Sep;59(3):740–742. doi: 10.1128/jvi.59.3.740-742.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Yuen L. K., Consigli R. A. Generation of capsids from unstable polyoma virions. J Virol. 1983 Sep;47(3):620–625. doi: 10.1128/jvi.47.3.620-625.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Yuen L. K., Consigli R. A. Identification and protein analysis of polyomavirus assembly intermediates from infected primary mouse embryo cells. Virology. 1985 Jul 15;144(1):127–138. doi: 10.1016/0042-6822(85)90311-3. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES