Identification of two DNA helicases UvrD and DinG as suppressors for lethality caused by mutant cspA mRNAs

Jihwan Hwang, Kangseok Lee, Sangita Phadtare, Masayori Inouye

Research output: Contribution to journalArticlepeer-review

4 Scopus citations


CspA is a major cold shock-inducible protein (70 aa), and its major role in the cold shock response was shown to be as an RNA chaperone destabilizing secondary structure of mRNAs at low temperature. Previously, we showed that the overexpression of mutant cspA containing premature non-sense codons at various positions led to stalled ribosomes on mutant cspA transcripts, ultimately leading to cell death. This lethality is primarily due to the highly translatable cspA 5′-UTR that recruits most of the ribosomes from other mRNAs, which are then stalled at the abnormal stop codon. This was called the 'LACE' effect. We show here that non-sense mutation even at the 67th position as well as substitutions of aromatic amino acid residues present on the RNA-binding surface of CspA protein to alanine caused the LACE effect by trapping a substantial amount of ribosomes on cspA mRNAs. In an attempt to identify a suppressor(s), which may help the cells to recover from the inhibitory LACE effect, genetic screening of an Escherichia coli genomic library was performed. We isolated suppressors that contained the genomic fragments encoding uvrD and dinG, respectively, whose gene products are ATP-dependent DNA helicases. The nucleic acid-binding and ATPase activities of these two helicases were found to be essential for their suppression activity. This genomic screening offers an approach to shed light on the mechanistic of 5′-UTR of cspA mRNA and novel roles of E. coli helicases that function in DNA repair.

Original languageEnglish (US)
Pages (from-to)135-146
Number of pages12
JournalJournal of Molecular Microbiology and Biotechnology
Issue number3
StatePublished - Aug 2012
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Microbiology
  • Applied Microbiology and Biotechnology
  • Molecular Biology


Dive into the research topics of 'Identification of two DNA helicases UvrD and DinG as suppressors for lethality caused by mutant cspA mRNAs'. Together they form a unique fingerprint.

Cite this