5th International AIDS Conference Montreal, Quebec, Canada — Jun 4-9, 1989

Int Conf AIDS 1989 Jun 4-9; 5:669 (abstract no. C.666)

Ferris A, Hizi A, Pichuantes S, Babe L, Craik C, Showalter SD, Hughes SH; BRI-Basic Research Program, Frederick, Maryland, USA

OBJECTIVE: We wished to test the ability of viral and non-viral proteases to convert purified 66-Kd reverse transcriptase (RT) to the 51-Kd form.

METHODS: The 66-Kd form of HIV-1 RT and the viral protease were separately purified to homogeneity from two recombinant strains of E. coli. Proteolytic cleavage sites on HIV-1 RT were mapped using a battery of monoclonal antibodies that recognize known positions in HIV-1 RT.

RESULTS: Using a viral protease produced in E. coli we have been able to reproduce the specific conversion of the 66-Kd form to the 51-Kd form in a reaction that contains only HIV-1 RT and the protease. However, this poses the question of whether the specificity of cleavage resides in the protease or in the structure of the properly folded 66-Kd form of the RT. We have found that certain other proteases produce cleavage products closely related to the 51-Kd protein produced by the viral protease.

CONCLUSIONS: Of the tested proteases, the viral protease most closely reproduces the cleavage event seen in virions; however, unrelated proteases make similar cleavages suggesting that the structure of the 66-Kd form of the RT plays a significant role in defining the site of protease cleavage.

Keywords: AEGIS, HIV-1 Reverse Transcriptase, RNA-Directed DNA Polymerase, HIV Protease, HIV-1, Endopeptidases, Escherichia coli, virology, ICA5

1989-06-04
C666

Copyright © 1989 - International AIDS Society (IAS). Reproduction of this abstract (other than one copy for personal reference) must be cleared through the IAS.