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HIV-1 Rev promotes the nuclear export of unspliced and singly spliced RNAs in a mammalian cell-free export system.

Publication ,  Journal Article
Huffman, KM; Arrigo, SJ; Schmidt, MG
Published in: J Biomed Sci
1999

Rev has been shown to promote the export of HIV-1 RNAs from Xenopus oocyte nuclei, but a system to examine the direct effect of Rev on HIV-1 RNA export in mammalian somatic cells does not exist. In this report, the development of a cell-free RNA export system using COS cells is described. This system is capable of examining the movement of RNA from nuclei of COS cells transfected with an HIV-1 proviral construct into reconstituted cytosol from nontransfected cells. A reproducible preparation of nuclei free of residual cytoplasmic RNA is demonstrated. Export of RNA from these nuclei into reconstituted cell-free extracts was saturable and dependent on temperature and energy. Further validation of the system was obtained by confirming that the nuclear export of HIV-1-unspliced and partially spliced RNAs was dependent upon the expression of HIV-1 Rev and that the presence of Rev appeared to decrease the export of an HIV-1-spliced RNA. The system was also able to demonstrate that Rev did not appear to significantly enhance the export of an HIV-1 protease-containing RNA that has been shown to be dependent upon Rev for maximal expression. Consequently, the system appears useful for the examination of parameters of nuclear export of HIV-1 and cellular RNAs.

Duke Scholars

Published In

J Biomed Sci

DOI

ISSN

1021-7770

Publication Date

1999

Volume

6

Issue

3

Start / End Page

194 / 205

Location

England

Related Subject Headings

  • rev Gene Products, Human Immunodeficiency Virus
  • Transfection
  • Temperature
  • RNA, Viral
  • RNA Splicing
  • RNA
  • HIV-1
  • Gene Products, rev
  • Energy Metabolism
  • DNA Primers
 

Citation

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MLA
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Huffman, K. M., Arrigo, S. J., & Schmidt, M. G. (1999). HIV-1 Rev promotes the nuclear export of unspliced and singly spliced RNAs in a mammalian cell-free export system. J Biomed Sci, 6(3), 194–205. https://doi.org/10.1007/BF02255903
Huffman, K. M., S. J. Arrigo, and M. G. Schmidt. “HIV-1 Rev promotes the nuclear export of unspliced and singly spliced RNAs in a mammalian cell-free export system.J Biomed Sci 6, no. 3 (1999): 194–205. https://doi.org/10.1007/BF02255903.
Huffman, K. M., et al. “HIV-1 Rev promotes the nuclear export of unspliced and singly spliced RNAs in a mammalian cell-free export system.J Biomed Sci, vol. 6, no. 3, 1999, pp. 194–205. Pubmed, doi:10.1007/BF02255903.
Journal cover image

Published In

J Biomed Sci

DOI

ISSN

1021-7770

Publication Date

1999

Volume

6

Issue

3

Start / End Page

194 / 205

Location

England

Related Subject Headings

  • rev Gene Products, Human Immunodeficiency Virus
  • Transfection
  • Temperature
  • RNA, Viral
  • RNA Splicing
  • RNA
  • HIV-1
  • Gene Products, rev
  • Energy Metabolism
  • DNA Primers