Effect of temperature on in vivo protein synthetic capacity in Escherichia coli.
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 degreesC. 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 degreesC. Unlike the growth rate, however, the elongation rate continues to increase up to 44 degreesC at the same rate as between 25 and 37 degreesC. 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 degreesC), 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.
Farewell
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Department of Microbiology and Immunology, University of Michigan, Ann Arbor, Michigan, USA.
Neidhardt
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FC
D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D004790:Enzyme Induction; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D010441:Peptide Chain Elongation, Translational; D012335:RNA, Ribosomal / Q000378:metabolism; D012269:Ribosomal Proteins / Q000378:metabolism; D013696:Temperature; D001616:beta-Galactosidase / Q000096:biosynthesis
D001426:Bacterial Proteins; D012335:RNA, Ribosomal; D012269:Ribosomal Proteins; D001616:beta-Galactosidase