MAMMALIAN NONSENSE CODONS CAN BE CIS EFFECTORS OF NUCLEAR MESSENGER-RNA HALF-LIFE

被引:126
作者
BELGRADER, P [1 ]
CHENG, J [1 ]
ZHOU, XB [1 ]
STEPHENSON, LS [1 ]
MAQUAT, LE [1 ]
机构
[1] ROSWELL PK CANC INST,DEPT HUMAN GENET,BUFFALO,NY 14263
关键词
D O I
10.1128/MCB.14.12.8219
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:8219 / 8228
页数:10
相关论文
共 40 条
[1]   ROUS-SARCOMA VIRUS-RNA STABILITY REQUIRES AN OPEN READING FRAME IN THE GAG GENE AND SEQUENCES DOWNSTREAM OF THE GAG-POL JUNCTION [J].
BARKER, GF ;
BEEMON, K .
MOLECULAR AND CELLULAR BIOLOGY, 1994, 14 (03) :1986-1996
[2]   NONSENSE CODONS WITHIN THE ROUS-SARCOMA VIRUS GAG GENE DECREASE THE STABILITY OF UNSPLICED VIRAL-RNA [J].
BARKER, GF ;
BEEMON, K .
MOLECULAR AND CELLULAR BIOLOGY, 1991, 11 (05) :2760-2768
[3]   BETA-GLOBIN NONSENSE MUTATION - DEFICIENT ACCUMULATION OF MESSENGER-RNA OCCURS DESPITE NORMAL CYTOPLASMIC STABILITY [J].
BASERGA, SJ ;
BENZ, EJ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (07) :2935-2939
[4]   CONSEQUENCES OF FRAMESHIFT MUTATIONS AT THE IMMUNOGLOBULIN HEAVY-CHAIN LOCUS OF THE MOUSE [J].
BAUMANN, B ;
POTASH, MJ ;
KOHLER, G .
EMBO JOURNAL, 1985, 4 (02) :351-359
[5]  
BAUMANN H, 1988, MOL CELL BIOL, V8, P42
[6]   MECHANISMS OF MESSENGER-RNA DECAY IN BACTERIA - A PERSPECTIVE [J].
BELASCO, JG ;
HIGGINS, CF .
GENE, 1988, 72 (1-2) :15-23
[7]   NONSENSE BUT NOT MISSENSE MUTATIONS CAN DECREASE THE ABUNDANCE OF NUCLEAR MESSENGER-RNA FOR THE MOUSE MAJOR URINARY PROTEIN, WHILE BOTH TYPES OF MUTATIONS CAN FACILITATE EXON SKIPPING [J].
BELGRADER, P ;
MAQUAT, LE .
MOLECULAR AND CELLULAR BIOLOGY, 1994, 14 (09) :6326-6336
[8]   EVIDENCE TO IMPLICATE TRANSLATION BY RIBOSOMES IN THE MECHANISM BY WHICH NONSENSE CODONS REDUCE THE NUCLEAR-LEVEL OF HUMAN TRIOSEPHOSPHATE ISOMERASE MESSENGER-RNA [J].
BELGRADER, P ;
CHENG, J ;
MAQUAT, LE .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (02) :482-486
[9]  
CHENG J, 1990, Molecular and Cellular Biology, V10, P5215
[10]   INTRONS ARE CIS EFFECTORS OF THE NONSENSE-CODON-MEDIATED REDUCTION IN NUCLEAR MESSENGER-RNA ABUNDANCE [J].
CHENG, J ;
BELGRADER, P ;
ZHOU, XB ;
MAQUAT, LE .
MOLECULAR AND CELLULAR BIOLOGY, 1994, 14 (09) :6317-6325