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Papers In Press, published online ahead of print December 12, 2001
Department of Biochemistry, UMDNJ-Robert Wood Johnson Medical School, Piscataway, New Jersey 08854
Corresponding Author: xiabi{at}umdnj.edu
The 5’-end region of the cspA mRNA contains a Cold Box sequence conserved among several cold-shock mRNAs. This region forms a stable stem-loop structure followed by an AU-rich sequence. Here we show that the Cold Box region is essential for the normal scale of cspA mRNA induction after cold shock, as a deletion of the stem-loop significantly destabilizes the mRNA and reduces the cold shock-induced cspA mRNA amount by approximately 50%. The AU-rich track, however, slightly destabilizes the mRNA. The integrity of the stem is essential for the stabilizing function, while that of the loop sequence is less important. Overexpression of a mutant cspA mRNA devoid of both the AUG initiation codon and the coding sequence results in a severe growth inhibition at low temperature along with a derepression of the chromosomal cspA expression. Further, the overexpressed RNA is stably associated with the 30S and 70S ribosomes. Our results demonstrate that the AUG initiation codon and the coding region containing the downstream box are not required for the cspA mRNA to bind ribosomes and that the 5’-UTR by itself has a remarkable affinity to ribosomes at low temperature.
J. Biol. Chem, 10.1074/jbc.M109700200
Submitted on October 8, 2001
Revised on December 12, 2001
Accepted on December 12, 2001
The Cold Box stem-loop proximal to the 5’-end of the Escherichia coli cspA gene stabilizes its mRNA at low temperature
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