Role of Escherichia coli cspA promoter sequences and adaptation of translational apparatus in the cold shock response.
A shift in growth temperature from 37 degrees C to 15 degrees 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 posttranscriptional 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 degrees 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 degrees 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 posttranscriptional controls, such as alterations in mRNA stability and the translational apparatus.
Goldenberg
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D
Department of Molecular Genetics and Biotechnology, Hebrew University-Hadassah Medical School, Jerusalem, Israel.
Azar
I
I
Oppenheim
A B
AB
Brandi
A
A
Pon
C L
CL
Gualerzi
C O
CO
D000222:Adaptation, Physiological; D001426:Bacterial Proteins / Q000235:genetics*; D003080:Cold Temperature; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D016296:Mutagenesis; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D011499:Protein Processing, Post-Translational
D001426:Bacterial Proteins; C062260:cold shock protein CS7.4, Bacteria