- 作者: Kim M. Huffman Salvatore J. Arrigo Michael G. Schmidt
- 作者服務機構: Department of Microbiology and Immunology, Medical University of South carolina, Charleston, S.C., USA
- 中文摘要: --
- 英文摘要: Rev has been shown to promote the export of HIV-1RNAs from Xenopus oocyte nuclei, but a system toexamine the direct effect of Rev on HIV-1 RNA export inmammalian somatic cells does not exist. In this report,the development of a cell-free RNA export system usingCOS cells is described. This system is capable of examin-ing the movement of RNA from nuclei of COS cells trans-fected with an HIV-1 proviral construct into reconstitutedcytosol from nontransfected cells. A reproducible prepa-ration of nuclei free of residual cytoplasmic RNA is dem-onstrated. Export of RNA from these nuclei into reconsti-tuted cell-free extracts was saturable and dependent ontemperature and energy. Further validation of the sys-tem was obtained by confirming that the nuclear exportof HIV-1-unspliced and partially spliced RNAs was de-pendent upon the expression of HIV-1 Rev and that thepresence of Rev appeared to decrease the export of anHIV-1-spliced RNA. The system was also able to demon-strate that Rev did not appear to significantly enhancethe export of an HIV-1 protease-containing RNA that hasbeen shown to be dependent upon Rev for maximalexpression. Consequently, the system appears useful forthe examination of parameters of nuclear export of HIV-1and cellular RNAs.
- 中文關鍵字: --
- 英文關鍵字: HIV-1 Rev. Nuclear export. cis acting repressor sequences. HIV-1 protease