PMID: 9451455Feb 6, 1998Paper

Regulation of protein synthesis by minigene expression

Biochimie
J HernándezGabriel Guarneros

Abstract

Peptidyl-tRNA hydrolase (Pth), an enzyme essential for Escherichia coli viability, scavenges peptidyl-tRNA released during abortive polypeptide chain elongation. Bacterial strains of E coli partially defective in Pth activity are unable to maintain bacteriophage lambda growth. Phage mutations that overcome the bacterial defect have been located to several regions in the lambda genome named bar. Plasmid constructs expressing just the bar region are toxic and cause a general arrest of protein synthesis in Pth-defective cells. Inspection of the nucleotide sequence from two bar regions reveals the short coding sequence AUG AUA Stop, spaced by an AT-rich segment from a Shine Dalgarno-like sequence (S-D). These sequences have been named minigenes. Base changes altering the putative S-D, the two sense codons, or the stop codon have been found to reduce Bar-toxicity. Transcripts containing bar function as mRNA. Upon expression in pth mutants, wild-type (bar+) transcripts are found associated with ribosomes. In addition, bar+ RNA forms ternary complexes with the 30S ribosomal subunit and the initiator tRNA and can be released upon run-off translation in the same way as an authentic mRNA. A cell free system for protein synthesis reproduc...Continue Reading

References

Feb 1, 1990·Journal of Bacteriology·P GuzmanG Guarneros
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Citations

Mar 23, 2004·Gene·Norma-Angélica OviedoGabriel Guarneros
Jan 3, 2001·Molecular Microbiology·J G ValadezG Guarneros
Jun 3, 2000·The EMBO Journal·V Heurgué-HamardM Ehrenberg
Feb 18, 2006·Journal of Bacteriology·Ming GongCharles Yanofsky
Jan 1, 1997·Annual Review of Biochemistry·N Sarkar
Jun 6, 2003·Molecular Microbiology·Dale LindsleyGabriel Guarneros
Nov 13, 2002·Microbiology·L Rogelio Cruz-VeraGabriel Guarneros
Jan 29, 2002·Protein Expression and Purification·Paul D BoninLaurence A Erickson
Feb 1, 2014·Protein Expression and Purification·Kasey Taylor-CreelRobert L McFeeters
Feb 27, 1999·Journal of Bacteriology·T TensonA S Mankin
Feb 21, 2006·Biochimie·Luis Delgado-OlivaresJavier Hernandez-Sanchez

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