Influences on gene expression in vivo by a Shine-Dalgarno sequence.

Jin, HainingStockholms universitet,Institutionen för genetik, mikrobiologi och toxikologi (författare)

The Shine-Dalgarno (SD+: 5'-AAGGAGG-3') sequence anchors the mRNA by basepairing to the 16S rRNA in the small ribosomal subunit during translation initiation. We have here comparedhow an SD+ sequence influences gene expression, if located upstream or downstream of an initiation codon.The positive effect of an upstream SD+ is confirmed. A downstream SD+ gives decreased gene expression.This effect is also valid for appropriately modified natural Escherichia coli genes. If an SD+ is placedbetween two potential initiation codons, initiation takes place predominantly at the second start site.The first start site is activated if the distance between this site and the downstream SD+ is enlargedand/or if the second start site is weakened. Upstream initiation is eliminated if a stable stem-loopstructure is placed between this SD+ and the upstream start site. The results suggest that the two startsites compete for ribosomes that bind to an SD+ located between them. A minor positive contribution toupstream initiation resulting from 3' to 5' ribosomal diffusion along the mRNA is suggested. Analysisof the E. coli K12 genome suggests that the SD+ or SD-like sequences are systematically avoided in theearly coding region suggesting an evolutionary significance.

NATURVETENSKAP Biologi Biokemi och molekylärbiologi hsv//swe
NATURAL SCIENCES Biological Sciences Biochemistry and Molecular Biology hsv//eng
Base Sequence
Binding Sites
Codon; Initiator/genetics/metabolism
Escherichia coli/*genetics/metabolism
Gene Expression Regulation; Bacterial
Genes; Bacterial/genetics
Genes; Reporter
Molecular Sequence Data
Nucleic Acid Conformation
Protein Biosynthesis/*genetics
RNA; Bacterial/metabolism
RNA; Messenger/metabolism
RNA; Ribosomal; 16S/metabolism
RNA; Transfer; Amino Acyl/chemistry/metabolism
Ribosomes/*metabolism

Zhao, QingStockholms universitet,Institutionen för genetik, mikrobiologi och toxikologi (författare)
Gonzalez de Valdivia, Ernesto IStockholms universitet,Institutionen för genetik, mikrobiologi och toxikologi (författare)
Ardell, David HUppsala universitet,Centrum för bioinformatik,Ardell (författare)
StenstrÃ¶m, MagnusStockholms universitet,Institutionen för genetik, mikrobiologi och toxikologi (författare)
Isaksson, Leif AStockholms universitet,Institutionen för genetik, mikrobiologi och toxikologi (författare)
Stockholms universitetInstitutionen för genetik, mikrobiologi och toxikologi (creator_code:org_t)

Av författaren/redakt...: Jin, Haining; Zhao, Qing; Gonzalez de Vald ...; Ardell, David H; StenstrÃ¶m, Magn ...; Isaksson, Leif A

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