Growth-rate-dependent regulation of 6-phosphogluconate dehydrogenase level mediated by an anti-Shine-Dalgarno sequence located within the Escherichia coli gnd structural gene.

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Previous work has shown that in Escherichia coli K-12 growth-rate-dependent regulation of expression of 6-phosphogluconate dehydrogenase, encoded by the gnd gene, occurs at the posttranscriptional level and is mediated by a negative control element that lies deep in the coding sequence, somewhere between codons 48 and 118. Deletion analysis of a growth-rate-regulated gnd-lacZ translational fusion showed that the element is the segment of gnd mRNA between codons 67 and 78 that is complementary to an extensive portion of the gnd ribosome-binding site, including its Shine-Dalgarno sequence. The boundaries of the element were further defined by the cloning of a synthetic "internal complementary sequence." The core internal complementary sequence element effected growth-rate-dependent regulation when placed at several sites between codon 40 and codon 69, but it severely reduced gene expression when moved to codon 13. The effect on regulation of single and double mutations introduced into the element by site-directed mutagenesis correlated with the ability of the respective mRNAs to fold into secondary structures that sequester the ribosome-binding site. Thus the gnd gene's internal regulatory element appears to function as a cis-acting antisense RNA.

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