PMID- 12950926
OWN - NLM
STAT- MEDLINE
DCOM- 20031211
LR  - 20190725
IS  - 0950-382X (Print)
IS  - 0950-382X (Linking)
VI  - 49
IP  - 6
DP  - 2003 Sep
TI  - Tetracycline-aptamer-mediated translational regulation in yeast.
PG  - 1627-37
AB  - We describe post-transcriptional gene regulation in yeast based on direct 
      RNA-ligand interaction. Tetracycline-dependent translational regulation could be 
      imposed via specific aptamers inserted at two different positions in the 5' 
      untranslated region (5'UTR). Translation in vivo was suppressed up to ninefold 
      upon addition of tetracycline. Repression via an aptamer located near the start 
      codon (cap-distal) in the 5'UTR was more effective than repression via a 
      cap-proximal position. On the other hand, suppression in a cell-free system 
      reached maximally 50-fold and was most effective via a cap-proximal aptamer. 
      Examination of the kinetics of tetracycline-dependent translational inhibition in 
      vitro revealed that preincubation of tetracycline and mRNA before starting 
      translation led not only to the fastest onset of inhibition but also the most 
      effective repression. The differences between the behaviour of the regulatory 
      system in vivo and in vitro are likely to be related to distinct properties of 
      mRNP structure and mRNA accessibility in intact cells as opposed to 
      cell-extracts. Tetracycline-dependent regulation was also observed after 
      insertion of an uORF sequence upstream of the aptamer, indicating that our system 
      also targets reinitiating ribosomes. Polysomal gradient analyses provided insight 
      into the mechanism of regulation. Cap-proximal insertion inhibits binding of the 
      43S complex to the cap structure whereas start-codon-proximal aptamers interfere 
      with formation of the 80S ribosome, probably by blocking the scanning 
      preinitiation complex.
FAU - Hanson, Shane
AU  - Hanson S
AD  - Lehrstuhl fur Mikrobiologie, Friedrich-Alexander Universitat Erlangen-Nurnberg, 
      Staudtstrasse 5, 91058 Erlangen, Germany.
FAU - Berthelot, Karine
AU  - Berthelot K
FAU - Fink, Barbara
AU  - Fink B
FAU - McCarthy, John E G
AU  - McCarthy JE
FAU - Suess, Beatrix
AU  - Suess B
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Mol Microbiol
JT  - Molecular microbiology
JID - 8712028
RN  - 0 (5' Untranslated Regions)
RN  - 0 (Luminescent Proteins)
RN  - 0 (Oligoribonucleotides)
RN  - 0 (RNA, Fungal)
RN  - 0 (RNA, Messenger)
RN  - F8VB5M810T (Tetracycline)
SB  - IM
MH  - 5' Untranslated Regions/genetics
MH  - Base Sequence
MH  - Blotting, Northern
MH  - Gene Expression Regulation, Fungal/*drug effects
MH  - Genes, Reporter
MH  - Luminescent Proteins/metabolism
MH  - Molecular Sequence Data
MH  - Oligoribonucleotides/genetics/metabolism
MH  - Plasmids
MH  - Polyribosomes/metabolism
MH  - Protein Biosynthesis/*drug effects
MH  - RNA, Fungal/genetics/metabolism
MH  - RNA, Messenger/genetics/metabolism
MH  - Saccharomyces cerevisiae/*drug effects/*genetics/metabolism
MH  - Tetracycline/*metabolism/pharmacology
MH  - Transformation, Genetic
EDAT- 2003/09/03 05:00
MHDA- 2003/12/12 05:00
CRDT- 2003/09/03 05:00
PHST- 2003/09/03 05:00 [pubmed]
PHST- 2003/12/12 05:00 [medline]
PHST- 2003/09/03 05:00 [entrez]
AID - 3656 [pii]
AID - 10.1046/j.1365-2958.2003.03656.x [doi]
PST - ppublish
SO  - Mol Microbiol. 2003 Sep;49(6):1627-37. doi: 10.1046/j.1365-2958.2003.03656.x.