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Journal of Bacteriology, March 2000, p. 1523-1528, Vol. 182, No. 6
Departamento de Genética y
Biología Molecular, Centro de Investigación y de
Estudios Avanzados del IPN, México
City,1 and Departamento de
Genética Molecular, Centro de Investigación sobre
Fijación de Nitrógeno, UNAM,
Cuernavaca,2 Mexico
Received 18 August 1999/Accepted 21 December 1999
The gene pth, encoding peptidyl-tRNA hydrolase (Pth),
is essential for protein synthesis and viability of Escherichia
coli. Two pth mutants have been studied in depth: a
pth(Ts) mutant isolated as temperature sensitive
and a pth(rap) mutant selected as nonpermissive for
bacteriophage
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Molecular Basis for the Temperature Sensitivity
of Escherichia coli pth(Ts)
vegetative growth. Here we show that each mutant
protein is defective in a different way. The Pth(Ts) protein was very
unstable in vivo, both at 43°C and at permissive temperatures, but its specific activity was comparable to that of the wild-type enzyme, Pth(wt). Conversely, the mutant Pth(rap) protein had
the same stability as Pth(wt), but its specific activity was
low. The thermosensitivity of the pth(Ts) mutant,
presumably, ensues after Pth(Ts) protein levels are reduced at 43°C.
Conditions that increased the cellular Pth(Ts) concentration, a rise in
gene copy number or diminished protein degradation, allowed cell growth at a nonpermissive temperature. Antibiotic-mediated inhibition of mRNA
and protein synthesis, but not of peptidyl-tRNA drop-off, reduced
pth(Ts) cell viability even at a permissive
temperature. Based on these results, we suggest that Pth(Ts) protein,
being unstable in vivo, supports cell viability only if its
concentration is maintained above a threshold that allows general
protein synthesis.
*
Corresponding author. Mailing address: Departamento de
Genética y Biología Molecular, Centro de
Investigación y de Estudios Avanzados del IPN, Apartado Postal
14-740, México D.F. 07000, Mexico. Phone: (52-5)7477000, ext.
5340. Fax: (52-5)7477100. E-mail: guarnero{at}gene.cinvestav.mx.
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