Hostname: page-component-848d4c4894-wzw2p Total loading time: 0 Render date: 2024-05-01T09:37:07.820Z Has data issue: false hasContentIssue false

Replacement of the yeast TRP4 3′ untranslated region by a hammerhead ribozyme results in a stable and efficiently exported mRNA that lacks a poly(A) tail

Published online by Cambridge University Press:  24 April 2002

KATRIN DÜVEL
Affiliation:
Institute of Microbiology and Genetics, Georg-August-University, D-37077 Göttingen, Germany Present address: Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544, USA.
OLIVER VALERIUS
Affiliation:
Institute of Microbiology and Genetics, Georg-August-University, D-37077 Göttingen, Germany
DAVID A. MANGUS
Affiliation:
Department of Molecular Genetics and Microbiology, University of Massachusetts Medical School, Worcester, Massachusetts 01655–0122, USA
ALLAN JACOBSON
Affiliation:
Department of Molecular Genetics and Microbiology, University of Massachusetts Medical School, Worcester, Massachusetts 01655–0122, USA
GERHARD H. BRAUS
Affiliation:
Institute of Microbiology and Genetics, Georg-August-University, D-37077 Göttingen, Germany
Get access

Abstract

The mRNA poly(A) tail serves different purposes, including the facilitation of nuclear export, mRNA stabilization, efficient translation, and, finally, specific degradation. The posttranscriptional addition of a poly(A) tail depends on sequence motifs in the 3′ untranslated region (3′ UTR) of the mRNA and a complex trans-acting protein machinery. In this study, we have replaced the 3′ UTR of the yeast TRP4 gene with sequences encoding a hammerhead ribozyme that efficiently cleaves itself in vivo. Expression of the TRP4-ribozyme allele resulted in the accumulation of a nonpolyadenylated mRNA. Cells expressing the TRP4-ribozyme mRNA showed a reduced growth rate due to a reduction in Trp4p enzyme activity. The reduction in enzyme activity was not caused by inefficient mRNA export from the nucleus or mRNA destabilization. Rather, analyses of mRNA association with polyribosomes indicate that translation of the ribozyme-containing mRNA is impaired. This translational defect allows sufficient synthesis of Trp4p to support growth of trp4 cells, but is, nevertheless, of such magnitude as to activate the general control network of amino acid biosynthesis.

Type
Research Article
Copyright
© 2002 RNA Society

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)