A NOVEL TRANSFORMING GROWTH FACTOR-BETA(1) RESPONSIVE CYTOPLASMIC TRANS-ACTING FACTOR BINDS SELECTIVELY TO THE 3'-UNTRANSLATED REGION OF MAMMALIAN RIBONUCLEOTIDE REDUCTASE-R2 MESSENGER-RNA - ROLE IN MESSAGE STABILITY

被引:42
作者
AMARA, FM
CHEN, FY
WRIGHT, JA
机构
[1] UNIV MANITOBA, MANITOBA INST CELL BIOL, 100 OLIVIA ST, WINNIPEG R3E 0V9, MANITOBA, CANADA
[2] UNIV MANITOBA, DEPT BIOCHEM & MOLEC BIOL, WINNIPEG R3E 0V9, MANITOBA, CANADA
基金
加拿大自然科学与工程研究理事会;
关键词
D O I
10.1093/nar/21.20.4803
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
Ribonucleotide reductase is a highly regulated enzyme that provides the four deoxyribonucleotides required for DNA synthesis. Our studies showed that TGF-beta1 treatment of BALB/c 3T3 mouse fibroblasts markedly elevated ribonucleotide reductase R2 mRNA levels, and also increased the half-life of R2 message by 4-fold from 1.5 h in untreated cells to 6 h in treated cells. We describe a novel 75 Kd sequence-specific cytoplasmic factor (p75) that binds selectively to a 83-nucleotide 3'-untranslated region of R2 mRNA and did not bind to the 5'UTR, the coding region of the R2 message or to the 3'UTRs of other mRNAs (from c-myc, GM-CSF and the iron responsive element from the transferrin receptor mRNA), or to the homopolymer poly(A) sequence. p75-RNA binding activity, which requires new protein synthesis, is not present in untreated cells, but is induced following TGF-beta1 stimulation. The in vivo kinetics of appearance of p75 binding activity paralleled the accumulation of R2 mRNA. Insertion of the 3'-untranslated region into the chloramphenicol acetyltransferase (CAT) message confers TGF-beta1 induced stability of RNA in stably transfected cells, while the same insert carrying a deletion of the 83-nucleotide fragment had little affect on RNA levels. Furthermore, in vitro decay reactions that contained the 83-nucleotide RNA or deletion of this fragment caused a significant decrease in TGF-beta1 stabilization of R2 message. A model is presented of R2 message regulation in which TGF-beta1 mediated stabilization of R2 message involves a specific interaction of a p75-trans-acting factor with a cis-element(s) stability determinant within the 83-nucleotide sequence which is linked to a reduction in the rate of R2 mRNA degradation.
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页码:4803 / 4809
页数:7
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