1887

Abstract

Aspartic proteinases from human imunodeficiency virus type 1 (HIV-1) and avian myeloblastosis virus (AMV) were found to interfere with microtubule assembly. Preincubation of the proteinases with purified brain microtubule proteins (tubulin and microtubule-associated proteins) at low ionic strength (pH 6·8), completely inhibited microtubule assembly. Analysis of microtubule proteins after incubation with proteinase showed no effect on tubulin but extensive cleavage of the microtubule-associated proteins 1 and 2 was observed. The digestion by the two proteinases differed. In the presence of HIV-1 proteinase, a fragment with an of approximately 300000 appeared, as well as at least three other new fragments, with values of 188000, 124000 and 73000. In the presence of AMV proteinase, the microtubule-associated proteins were extensively digested to many small fragments. The extending microtubule-associated proteins normally seen by electron microscopy on the microtubule surface disappeared after treatment with AMV proteinase. Our results show that retroviral proteinases are not restricted to cleavage of viral polyproteins It is suggested that proteolysis of microtubular proteins by viral proteinases is an important step in viral pathogenicity and that it may be part of a mechanism causing degenerative effects in infected cells.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-71-9-1985
1990-09-01
2024-12-09
Loading full text...

Full text loading...

/deliver/fulltext/jgv/71/9/JV0710091985.html?itemId=/content/journal/jgv/10.1099/0022-1317-71-9-1985&mimeType=html&fmt=ahah

References

  1. Blum H., Beier H., Gross H. J. 1987; Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis 8:93–99
    [Google Scholar]
  2. Darke P. L., Nutt R. F., Brady S. F., Garsky V. M., Ciccarone T. M., Leu C.-T., Lumma P. K., Freidinger R. M., Veber D. F., Sigal I. S. 1988; HIV-1 protease specificity of peptide cleavage is sufficient for processing of gag and pol polyproteins. Biochemical and Biophysical Research Communications 156:297–303
    [Google Scholar]
  3. de la Monte S. M., Ho D. D., Schooley R. T., Hirsch M. S., Richardson E. P. 1987; Subacute encephalomyelitis of AIDS and its relation to HTLV-III infection. Neurology 37:562–569
    [Google Scholar]
  4. Dittmar K., Moelling K. J. 1978; Biochemical properties of pl5- associated protease in an avian RNA tumor virus. Journal of Virology 28:106–118
    [Google Scholar]
  5. Dustin P. 1984 Microtubules Berlin: Springer Verlag;
    [Google Scholar]
  6. Hansen J., Billich S., Schulze T., Sukrow S., Moelling K. 1988; Partial purification and substrate analysis of bacterially expressed HIV protease by means of monoclonal antibody. EMBO Journal 7:1785–1791
    [Google Scholar]
  7. Joly J. C., Flynn G., Purich D. L. 1989; The microtubulebinding fragment of microtubule-associated protein-2. Location of the protease-accessible site and identification of an assembly- promoting peptide. Journal of Cell Biology 109:2289–2294
    [Google Scholar]
  8. Kotler M., Danho W., Katz R. A., Leis J., Skalka A. M. 1989; Avian retroviral protease and cellular aspartic proteases are distinguished by activities on peptide substrates. Journal of Biological Chemistry 264:3428–3435
    [Google Scholar]
  9. Kräusslich H.-G., Wimmer E. 1988; Viral proteinases. Annual Review of Biochemistry 57:701–754
    [Google Scholar]
  10. Kräusslich H.-G., Ingraham R. H., Skoog M. T., Wimmer E., Pallai P. V., Carter C. A. 1989; Activity of purified biosynthetic proteinase of human immunodeficiency virus on natural substrates and synthetic peptides. Proceedings of the National Academy of Sciences, U.S.A 86:807–811
    [Google Scholar]
  11. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  12. Lewis S. A., Wang D., Cowan N. J. 1988; Microtubule-associated protein MAP2 shares a microtubule binding motif with tau protein. Science 242:936–939
    [Google Scholar]
  13. Matus A., Green G. D. 1987; Age-related increase in a cathepsin D like protease that degrades brain microtubule-associated proteins. Biochemistry 26:8083–8086
    [Google Scholar]
  14. Peng C., Ho B. K., Chang T. W., Chang N. T. 1989; Role of human immunodeficiency virus type 1-specific protease in core protein maturation and viral infectivity. Journal of Virology 63:2550–2556
    [Google Scholar]
  15. Schägger H., von Jagow G. 1987; Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Analytical Biochemistry 166:369–379
    [Google Scholar]
  16. Schiff P. B., Horwitz S. B. 1981; Taxol assembles tubulin in the absence of exogenous guanosine 5′-triphosphate or microtubule-associated proteins. Biochemistry 20:3247–3252
    [Google Scholar]
  17. Serrano L., de la Torre J., Maccioni R. B., Avila J. 1984; Involvement of the carboxyl-terminal domain of tubulin in the regulation of its assembly. Proceedings of the National Academy of Sciences, U.S.A 81:5989–5993
    [Google Scholar]
  18. Sirianni M. C., Soddus S., Malorni W., Arancia G., Aiuti F. 1988; Mechanism of defective natural killer cell activity in patients with AIDS is associated with defective distribution of tubulin. Journal of Immunology 140:2565–2568
    [Google Scholar]
  19. Vallee R. B., Borisy G. G. 1977; Removal of the projections from cytoplasmic microtubules in vitro by digestion. Journal of Biological Chemistry 256:377–382
    [Google Scholar]
  20. Wallin M., Nordh J., Deinum J. 1986; Changes in the hydrodynamic properties of microtubules induced by taxol. Biochimica et biophysica acta 880:189–196
    [Google Scholar]
/content/journal/jgv/10.1099/0022-1317-71-9-1985
Loading
/content/journal/jgv/10.1099/0022-1317-71-9-1985
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error