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J. Biol. Chem., Vol. 277, Issue 8, 6005-6011, February 22, 2002
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,
From the Department of Biochemistry, University of Medicine and
Dentistry of New Jersey-Robert Wood Johnson Medical School, Piscataway,
New Jersey 08854
The 5'-end region of 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 because a deletion of the stem-loop significantly destabilizes
the mRNA and reduces the cold shock-induced cspA
mRNA amount by ~50%. The AU-rich track, however, slightly
destabilizes the mRNA. The integrity of the stem is essential for
the stabilizing function, whereas 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. Furthermore, the
overexpressed RNA is stably associated with the 30 S and 70 S
ribosomes. Our results demonstrate that the AUG initiation codon
and the coding region containing the downstream box are not required
for cspA mRNA to bind ribosomes and that the
5'-untranslated region by itself has a remarkable affinity to ribosomes
at low temperature.
Present address: Dept. of Cancer Biology, Dana-Farber Cancer
Institute and Harvard Medical School, 44 Binney St., Boston, MA 02115.
§
To whom correspondence should be addressed: Dept. of Biochemistry,
UMDNJ-Robert Wood Johnson Medical School, 675 Hoes Lane, Piscataway, NJ
08854. Tel.: 732-235-4115; Fax: 732-235-4559; E-mail: inouye@umdnj.edu.
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