Compositions and methods for regulating peptidyltransferase activity and uses thereof
The present invention provides compositions and means to identify compositions that increase −1 PRF programmed ribosomal frameshift (−1PRF) efficiencies and/or decrease peptidyltransferase activity in a cell, and thus directly affect viral replication or assembly of viral particles. Compositions identified in accordance with the invention specifically inhibit the interaction between ribosomal protein L41 and the ribosomes thereby resulting in decreased peptidyltransferase activity of the ribosomes. Decreases in peptidyltransferase activity have been associated with increased −1 PRF efficiencies, which in turn interfere with self assembly of −1PRF dependent viruses thereby interfering with virus propagation. Compositions in accordance with the invention are useful as antiviral therapeutics for treating a viral infection in a patient.
1. A method of identifying a composition which inhibits peptidyl transferase activity by inhibiting the interaction of L41 with ribosomes, comprising the steps of:
a) contacting a test composition with a cell, a cell extract, or purified cell components containing L41 and ribosomes, wherein the test composition is a small molecule or polypeptide;
b) detecting whether the test composition specifically inhibits the interaction of L41 with the ribosomes; and
c) determining whether the test composition inhibits peptidyl transferase activity of the ribosomes.
2. The method of claim 1 , wherein the cell or cell extract is from yeast.
3. The method of claim 1 , wherein the determining step comprises detecting a decrease in the rate of peptidylpuromycin formation by the ribosomes.
4. A method of identifying a composition which increases −1 programmed ribosomal frameshift (PRF) efficiencies by inhibiting the interaction of L41 with ribosomes comprising the steps of:
a) contacting a test composition with a cell, a cell extract, or purified cell components, containing L41 and ribosomes, wherein the test compound is a small molecule or polypeptide;
b) detecting whether the test composition inhibits the interaction of L41 with the ribosomes; and
c) determining whether the test composition increases −1 programmed ribosomal frameshift (PRF) efficiencies of the ribosomes.
5. The method of claim 4 , wherein the cell is a yeast cell.
6. The method of claim 4 , wherein the determining step comprises an assay of programmed ribosomal frameshifting.