A shift in growth temperature from 37° C to 15° C leads to a dramatic increase in the level of CspA, the major cold shock
protein of
Escherichia coli. To investigate the molecular basis of this induction, we considered the relevance of transcriptional and post-transcriptional
controls by analyzing the steady-state levels of transcripts and the expression of reporter genes in cells carrying a set
of
cspA promoter fragments of variable length fused to
lacZ or
cat genes. We demonstrate that: (i) the core
cspA promoter (from –40 to +16) responds to cold shock and a mutation at –36 increases the relative activity of the promoter at
low temperature by threefold; (ii) the sequences upstream of –40 have a positive effect on expression at 37° C, but no effect
on the cold shock response; (iii) by virtue of their influence on mRNA stability, the downstream sequences (from +81 to +165)
reduce expression at 37° C and increase the intensity of the cold shock response; (iv) mutations in the GCACATCA and CCAAT
motifs, present at +1/–4 and between the –10 and –35 elements, respectively, do not affect the cold shock response of the
cspA promoter; (v) following cold shock, a modification of the protein synthetic machinery takes place that allows preferential
translation of
cspA mRNA relative to the non-cold shock
cat and
lacZ mRNAs. The quantitatively modest transcriptional activation shown by the core promoter of
cspA following cold shock suggests that transcriptional activation can significantly contribute to cold shock induction only when
coupled to post-transcriptional controls, such as alterations in mRNA stability and of the translational apparatus.
Key words Cold shock - Core promoter activity - CCAAT sequence - Translational control - mRNA stability
Received: 20 February 1997 / Accepted: 12 June 1997