Mammalian nonsense codons can be cis effectors of nuclear mRNA half-life

Author:

Belgrader P1,Cheng J1,Zhou X1,Stephenson L S1,Maquat L E1

Affiliation:

1. Department of Human Genetics, Roswell Park Cancer Institute, Buffalo, New York 14263.

Abstract

Frameshift and nonsense mutations within the gene for human triosephosphate isomerase (TPI) that generate a nonsense codon within the first three-fourths of the protein coding region have been found to reduce the abundance of the product mRNA that copurifies with nuclei. The cellular process and location of the nonsense codon-mediated reduction have proven difficult to elucidate for technical reasons. We show here, using electron microscopy to judge the purity of isolated nuclei, that the previously established reduction to 25% of the normal mRNA level is evident for nuclei that are free of detectable cytoplasmic contamination. Therefore, the reduction is likely to be characteristic of bona fide nuclear RNA. Fully spliced nuclear mRNA is identified by Northern (RNA) blot hybridization and a reverse transcription-PCR assay as the species that undergoes decay in experiments that used the human c-fos promoter to elicit a burst and subsequent shutoff of TPI gene transcription upon the addition of serum to serum-deprived cells. Finally, the finding that deletion of a 5' splice site of the TPI gene results predominantly but not exclusively in the removal by splicing (i.e., skipping) of the upstream exon as a part of the flanking introns has been used to demonstrate that decay is specific to those mRNA products that maintain the nonsense codon. This result, together with our previous results that implicate translation by ribosomes and charged tRNAs in the decay mechanism, indicate that nonsense codon recognition takes place after splicing and triggers decay solely in cis. The possibility that decay takes place during the process of mRNA export from the nucleus to the cytoplasm is discussed.

Publisher

American Society for Microbiology

Subject

Cell Biology,Molecular Biology

Reference40 articles.

1. Nonsense codons within the Rous sarcoma virus gag gene decrease the stability of unspliced viral RNA;Barker G. F.;Mol. Cell. Biol.,1991

2. Rous sarcoma virus RNA stability requires an open reading frame in the gag gene and sequences downstream of the gag-pol junction;Barker G. F.;Mol. Cell. Biol.,1994

3. P-Globin nonsense mutation: deficient accumulation of mRNA occurs despite normal cytoplasmic stability;Baserga S. J.;Proc. Natl. Acad. Sci. USA,1992

4. Consequences of frameshift mutations at the immunoglobulin heavy chain locus of the mouse;Baumann B.;EMBO J.,1985

5. Hepatocyte-stimulating factor B2 interferon and interleukin-1 enhance expression of the rat (xl-acid glycoprotein gene via a distal upstream regulatory region;Baumann H.;Mol. Cell. Biol.,1988

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