Faculty Collaboration Database

The 5'-untranslated region of GM-CSF mRNA suppresses translational repression mediated by the 3' adenosine-uridine-rich element and the poly(A) tail. Nucleic Acids Res 1999 Sep 15;27(18):3660-6

Date

09/03/1999

Pubmed ID

10471734

Pubmed Central ID

PMC148620

DOI

10.1093/nar/27.18.3660

Scopus ID

2-s2.0-0033568389 (requires institutional sign-in at Scopus site)   12 Citations

Abstract

Granulocyte-macrophage colony stimulating factor (GM-CSF) mRNA levels are controlled post-transcriptionally by the 3'-untranslated region (UTR) adenosine-uridine-rich element (ARE). In untransformed, resting cells, the ARE targets GM-CSF mRNA for rapid degradation, thereby significantly suppressing protein expression. We used a rabbit reticulocyte lysate (RRL) cell-free system to examine translational regulation of GM-CSF expression. We uncoupled decay rates from rates of translation by programming the RRL with an excess of mRNAs. Capped, full-length, polyadenyl-ated human GM-CSF mRNA (full-length 5'-UTR AUUUA+A90) and an ARE-modified version (full-length 5'-UTR AUGUA+A90) produced identical amounts of protein. When the 5'-UTR was replaced with an irrelevant synthetic leader sequence (syn 5'-UTR), translation of syn 5'-UTR AUUUA+A90 mRNA was suppressed by >20-fold. Mutation of the ARE or removal of the poly(A) tail relieved this inhibition. Thus, in the absence of a native 5'-UTR, the ARE and poly(A) tail act in concert to block GM-CSF mRNA translation. Substitutions of different regions of the native 5'-UTR revealed that the entire sequence was essential in maintaining the highest rates of translation. However, shorter 10-12 nt contiguous 5'-UTR regions supported 50-60% of maximum translation. The 5'-UTR is highly conserved, suggesting similar regulation in multiple species and in these studies was the dominant element regulating GM-CSF mRNA translation, overriding the inhibitory effects of the ARE and the poly(A) tail.

Author List

Jarzembowski JA, Rajagopalan LE, Shin HC, Malter JS

Author

Jason A. Jarzembowski MD, PhD Sr Associate Dean, CEO CSG, Professor in the Pathology department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

5' Untranslated Regions
Adenosine
Base Sequence
Binding Sites
Conserved Sequence
Gene Silencing
Granulocyte-Macrophage Colony-Stimulating Factor
Humans
Kinetics
Mutation
Nucleic Acid Conformation
Poly A
Protein Biosynthesis
RNA, Messenger
Regulatory Sequences, Nucleic Acid
Temperature
Uridine