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Journal of Bacteriology, March 1999, p. 1617-1622, Vol. 181, No. 5
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Inhibition of Translation and Cell Growth by
Minigene Expression
Tanel
Tenson,1,2
Jesús Vega
Herrera,3
Patricia
Kloss,1
Gabriel
Guarneros,3 and
Alexander S.
Mankin1,*
Center for Pharmaceutical Biotechnology,
University of Illinois, Chicago, Illinois
606071; Estonian Biocenter, Tartu,
EE2400, Estonia2; and Departamento de
Genética y Biología Molecular, CINVESTAV-IPN, Mexico
City 07000, Mexico3
Received 31 August 1998/Accepted 26 December 1998
A random five-codon gene library was used to isolate minigenes
whose expression causes cell growth arrest. Eight different deleterious
minigenes were isolated, five of which had in-frame stop codons; the
predicted expressed peptides ranged in size from two to five amino
acids. Mutational analysis demonstrated that translation of the
inhibitory minigenes is essential for growth arrest. Pulse-labeling
experiments showed that expression of at least some of the selected
minigenes results in inhibition of cellular protein synthesis.
Expression of the deleterious minigenes in cells deficient in
peptidyl-tRNA hydrolase causes accumulation of families of
peptidyl-tRNAs corresponding to the last minigene codon; the inhibitory
action of minigene expression could be suppressed by overexpression of
the tRNA corresponding to the last sense codon in the minigene.
Experimental data are compatible with the model that the deleterious
effect of minigene expression is mediated by depletion of corresponding
pools of free tRNAs.
*
Corresponding author. Mailing address: Center for
Pharmaceutical Biotechnology
m/c 870, University of Illinois, 900 S. Ashland Ave., Chicago, IL 60607. Phone: (312) 413-1406. Fax: (312)
413-9303. E-mail: shura{at}uic.edu.
Journal of Bacteriology, March 1999, p. 1617-1622, Vol. 181, No. 5
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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