Zhang MJ, Dayton E, Dayton AI; International Conference on AIDS.
Int Conf AIDS. 1994 Aug 7-12; 10: 27 (abstract no. 400A).
LIR/NIAID/NIH, Bethesda, MD 20892.
OBJECTIVES: We are promoting a novel methodology to identify and clone cellular factors mechanistically involved in the Rev regulatory pathway. METHODS: We have formed Rev/RRE complexes in the presence of nuclear extracts to look for cooperative interactions. Ternary complexes formed by Rev, cellular factors and 32P-labeled RRE are stabilized by UV crosslinking and digested with ribonuclease to remove non-specific complexes. The complexes are then visualized by native gel electrophoresis. RESULTS: At concentrations of Rev and RRE which otherwise form no complexes, the addition of nuclear extracts induces the formation of large amounts of a broadly retarded complex. The degree of retardation is proportional to the amount of added extract. Complexes are not formed with mutant RREs which are defective for Rev binding or with anti-sense RRE. Data will be presented on the purification of factors which form these complexes and on their cell-type distribution. Data will also be presented on regions of the Rev protein and regions of the RRE required for complex formation. CONCLUSIONS: Nuclear extracts contain factors which cooperatively interact with Rev and the RRE.
Publication Types:
Keywords:
- Gene Products, rev
- HIV-1
- RNA, Antisense
Other ID:
UI: 102210571
From Meeting Abstracts