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Molecular and Cellular Biology, January 1999, p. 384-391, Vol. 19, No. 1
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Ribosomal Protein L3 Mutants Alter Translational Fidelity and Promote Rapid Loss of the Yeast Killer Virus

Stuart W. Peltz,1 Amy B. Hammell,2 Ying Cui,2 Jason Yasenchak,2 Lara Puljanowski,2 and Jonathan D. Dinman1,2,*

Department of Molecular Genetics and Microbiology and Graduate Program in Molecular Biosciences at UMDNJ/Rutgers Universities, Robert Wood Johnson Medical School-UMDNJ,2 and The Cancer Institute of New Jersey,1 Piscataway, New Jersey 08854

Received 23 July 1998/Returned for modification 3 September 1998/Accepted 28 September 1998

Programmed -1 ribosomal frameshifting is utilized by a number of RNA viruses as a means of ensuring the correct ratio of viral structural to enzymatic proteins available for viral particle assembly. Altering frameshifting efficiencies upsets this ratio, interfering with virus propagation. We have previously demonstrated that compounds that alter the kinetics of the peptidyl-transfer reaction affect programmed -1 ribosomal frameshift efficiencies and interfere with viral propagation in yeast. Here, the use of a genetic approach lends further support to the hypothesis that alterations affecting the ribosome's peptidyltransferase activity lead to changes in frameshifting efficiency and virus loss. Mutations in the RPL3 gene, which encodes a ribosomal protein located at the peptidyltransferase center, promote approximately three- to fourfold increases in programmed -1 ribosomal frameshift efficiencies and loss of the M1 killer virus of yeast. The mak8-1 allele of RPL3 contains two adjacent missense mutations which are predicted to structurally alter the Mak8-1p. Furthermore, a second allele that encodes the N-terminal 100 amino acids of L3 (called L3Delta ) exerts a trans-dominant effect on programmed -1 ribosomal frameshifting and killer virus maintenance. Taken together, these results support the hypothesis that alterations in the peptidyltransferase center affect programmed -1 ribosomal frameshifting.

* Corresponding author. Mailing address: Department of Molecular Genetics and Microbiology and Graduate Program in Molecular Biosciences at UMDNJ/Rutgers Universities, 675 Hoes Lane, Piscataway, NJ 08854. Phone: (732) 235-4670. Fax: (732) 235-5223. E-mail: dinmanjd{at}umdnj.edu.

Molecular and Cellular Biology, January 1999, p. 384-391, Vol. 19, No. 1
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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