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A codon window in mRNA downstream of the initiation codon where NGG codons give strongly reduced gene expression in Escherichia coli
Stockholm University, Faculty of Science, Department of Genetics, Microbiology and Toxicology.
2004 In: Nucleic Acids Reseach, Vol. 32, no 17, 5198-5205 p.Article in journal (Refereed) Published
Place, publisher, year, edition, pages
2004. Vol. 32, no 17, 5198-5205 p.
Identifiers
URN: urn:nbn:se:su:diva-25906OAI: oai:DiVA.org:su-25906DiVA: diva2:200781
Note
Part of urn:nbn:se:su:diva-985Available from: 2006-04-19 Created: 2006-04-19Bibliographically approved
In thesis
1. Studies on translation initiation and gene expression in Escherichia coli
Open this publication in new window or tab >>Studies on translation initiation and gene expression in Escherichia coli
2006 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

In prokaryotes, several mRNA sequences surrounding the initiation codon have been found to influence the translation process; these include the downstream region and its codon context, the Shine-Dalgarno sequence and the S1 ribosomal protein-binding site. In this thesis, the purpose has been to study the role of the downstream region and Shine-Dalgarno-like sequences on early translation elongation and gene expression in Escherichia coli.

The downstream region (DR) after the initiation codon (around five to seven codons), has an important role in the initiation of translation. We find that most of the codons which give very low gene expression at +2 (considering AUG as +1), reach 5 to 10 fold higher expression when those codons are positioned posteriori to +2, with the exception of the NGG codons. The NGG codons abort the translation process if located within the first five codons of the DR, due to peptidyl-tRNA drop-off. However, when the NGG codons are situated further down from the DR, the protein expression was increased at the same level of expression as in the presence of any other codon.

The Shine-Dalgarno (SD) is an important region of initiation in translation of bacteria. In spite of this, it has been found that Gram-negative bacteria could translate mRNAs with weak or non-functional SD, while the DR carries out a main role in the efficiency of translation. In addition, positions of SD and SD-like sequences are very important to direct initiation of translation in the choice between two possible initiation codons. A strong SD between two initiation sites will favor the second initiation site if it consists of a canonical start codon followed by a good DR.

The results suggest that the mRNA sequences surrounding the initiation codon: the downstream region and the Shine-Dalgarno and SD-like sequences, are very important contributors to the translation level and gene expression in Escherichia coli.

Place, publisher, year, edition, pages
Stockholm: Institutionen för genetik, mikrobiologi och toxikologi, 2006. 50 p.
Keyword
Downstream region, NGG codons, abortive translation, drop-off, Shine-Dalgarno sequence, gene expression and Escherichia coli
National Category
Natural Sciences
Identifiers
urn:nbn:se:su:diva-985 (URN)91-7155-214-6 (ISBN)
Public defence
2006-05-10, Nordenskiöldsalen, Geovetenskapens hus U, Svante Arrhenius väg 8 B, Stockholm, 13:00
Opponent
Supervisors
Available from: 2006-04-19 Created: 2006-04-19Bibliographically approved

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