PROGRAM

BN Seminar - Lars-Anders Carlson: 'In vitro reconstitution of selective HIV-1 RNA packaging by membrane-bound Gag assemblies'

Date:

Time:

14:00 hrs

Location:

A1.100 , TU Delft

 

The Gag protein of HIV-1 is the virus assembly engine and protein "endoskeleton" of the released virus particle. Gag selects and packages a dimeric, unspliced viral RNA in the context of a large excess of cytosolic human RNAs. As Gag assembles on the plasma membrane, the HIV-1 genome is enriched relative to cellular RNAs by an unknown mechanism. We used a minimal system consisting of purified RNAs, recombinant HIV-1 Gag and giant unilamellar vesicles to recapitulate the selective packaging of the 5’ untranslated region of the HIV-1 genome in the presence of excess competitor RNA. Mutations in the CA-CTD domain of Gag which subtly affect the self-assembly of Gag abrogated RNA selectivity. We further found that tRNA increases the RNA selectivity of assembling Gag.

The ability of HIV-1 Gag to selectively package its RNA genome and its self-assembly on membranes are thus interdependent on one another.

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