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Journal of Bacteriology, September 1998, p. 4704-4710, Vol. 180, No. 17
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Effect of Temperature on In Vivo Protein
Synthetic Capacity in Escherichia coli
Anne
Farewell* and
Frederick C.
Neidhardt
Department of Microbiology and Immunology,
University of Michigan, Ann Arbor, Michigan
Received 9 February 1998/Accepted 20 June 1998
In this report, we examine the effect of temperature on protein
synthesis. The rate of protein accumulation is determined by three
factors: the number of working ribosomes, the rate at which ribosomes
are working, and the rate of protein degradation. Measurements of
RNA/protein ratios and the levels of individual ribosomal
proteins and rRNA show that the cellular amount of ribosomal machinery in Escherichia coli is constant between 25 and
37°C. Within this range, in a given medium, temperature affects
ribosomal function the same as it affects overall growth. Two
independent methodologies show that the peptide chain elongation rate
increases as a function of temperature identically to
growth rate up to 37°C. Unlike the growth rate, however, the
elongation rate continues to increase up to 44°C at the same rate as
between 25 and 37°C. Our results show that the peptide elongation
rate is not rate limiting for growth at high temperature. Taking into
consideration the number of ribosomes per unit of cell mass, there is
an apparent excess of protein synthetic capacity in these cells,
indicating a dramatic increase in protein degradation at high
temperature. Temperature shift experiments show that peptide
chain elongation rate increases immediately, which supports a mechanism
of heat shock response induction in which an increase in unfolded,
newly translated protein induces this response. In addition, we find that at low temperature (15°C), cells contain a pool of
nontranslating ribosomes which do not contribute to cell growth,
supporting the idea that there is a defect in initiation at low
temperature.
*
Corresponding author. Present address: Dept. of
Microbiology, Lund University, Sölvegatan 12, 223 62 Lund,
Sweden. Phone: 46 46 222 0463. Fax: 46 46 15 78 39. E-mail:
anne.farewell{at}mikrbiol.lu.se.
Journal of Bacteriology, September 1998, p. 4704-4710, Vol. 180, No. 17
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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