The termination of protein synthesis in ribosomes is governed by termination (stop) codons in messenger RNAs and by polypeptide chain release factors (RFs). Although the primary structure of prokaryotic RFs and yeast mitochrondrial RF is established, that of the only known eukaryotic RF (eRF) remains obscure. Here we report the assignment of a family of tightly related proteins (designated eRF1) from lower and higher eukaryotes which are structurally and functionally similar to rabbit eRF. Two of these proteins, one from human and the other from Xenopus laevis, have been expressed in yeast and Escherichia coli, respectively, purified and shown to be active in the in vitro RF assay. The other protein of this family, sup45 (sup1) of Saccharomyces cerevisiae, is involved in omnipotent suppression during translation. The amino-acid sequence of the eRF1 family is highly conserved. We conclude that the eRF1 proteins are directly implicated in the termination of translation in eukaryotes.

The availability of full genome sequences has allowed the construction of microarrays, with which screening of the full genome for changes in gene expression is possible. This method can provide a wealth of information about biology at the level of gene expression and is a powerful method to identify genes and pathways involved in various processes. In this study, we report a detailed analysis of the full heat stress response in Drosophila melanogaster females, using whole genome gene expression arrays (Affymetrix Inc, Santa Clara, CA, USA). The study focuses on up- as well as downregulation of genes from just before and at 8 time points after an application of short heat hardening (36 degrees C for 1 hour). The expression changes were followed up to 64 hours after the heat stress, using 4 biological replicates. This study describes in detail the dramatic change in gene expression over time induced by a short-term heat treatment. We found both known stress responding genes and new candidate genes, and processes to be involved in the stress response. We identified 3 main groups of stress responsive genes that were early-upregulated, early-downregulated, and late-upregulated, respectively, among 1222 differentially expressed genes in the data set. Comparisons with stress sensitive genes identified by studies of responses to other types of stress allow the discussion of heat-specific and general stress responses in Drosophila. Several unexpected features were revealed by this analysis, which suggests that novel pathways and mechanisms are involved in the responses to heat stress and to stress in general. The majority of stress responsive genes identified in this and other studies were downregulated, and the degree of overlap among downregulated genes was relatively high, whereas genes responding by upregulation to heat and other stress factors were more specific to the stress applied or to the conditions of the particular study. As an expected exception, heat shock genes were generally found to be upregulated by stress in general.

1. Inducible heat‐shock proteins are synthesized when temperatures are increased to levels substantially above normal. The functional role of these proteins is well known at the cellular level. Today increasing interest has been directed towards the importance of heat‐shock proteins for resistance of whole organisms to high‐temperature stress and other environmental stressors. 2. Here the functional relationship between the heat‐shock protein, Hsp70, and thermal resistance in adult Drosophila melanogaster was examined by comparing thermal resistance, i.e. survival at 39 °C for 85 min, and levels of Hsp70 at various times elapsed (2, 4, 8, 16, 32 and 64 h) after thermotolerance was induced by short‐term acclimation/heat hardening at 37 °C for 55 min. 3. Levels of Hsp70 in both males and females were highest 2 h after heat hardening and declined with longer times elapsed. The rate of decrease initially was very fast but diminished with increasing time. After 32 h the level of Hsp70 approached the level in flies that were not hardened. Levels of Hsp70 in males exceeded that of females during the entire period. 4. Survival of both sexes increased with increasing time after heat hardening and reached an optimum between 8 and 32 h. Thereafter resistance decreased with longer times elapsed. Survival of females generally exceeded that of males except after 16 and 64 h. 5. Regression analysis applied to the data on Hsp70 levels revealed that the model describing these data could not explain the data for survival. Also, higher levels of Hsp70 in males compared with females were not associated with greater survival in males. However, statistical analysis on paired measurements of Hsp70 and survival revealed a positive association between Hsp70 level and survival at each time elapsed after induction of thermotolerance.

No abstract

The diadenosine oligophosphates (Ap n A) were discovered in the mid-sixties in the course of studies on aminoacyltRNA synthetases (aaRS). Now, more than 30 years later, about 300 papers have been published around these substances in attempt to decipher their role in cells. Recently, Ap n A have emerged as intracellular and extracellular signalling molecules implicated in the maintenance and regulation of vital cellular functions and become considered as second messengers. Great variety of physiological and pathological effects in mammalian cells was found to be associated with alterations of Ap n A levels (n from 2 to 6) and Ap Q A/Ap R A ratio. Cell differentiation and apoptosis have substantial and opposite effects on Ap Q A/Ap R A ratio in cultured cells. A human Ap Q A hydrolase, Fhit, appeared to be involved in protection of cells against tumourigenesis. Ap Q A is synthesised by mammalian u synthetase (TrpRS) which in contrast to most other aaRS is unable to synthesise Ap R A and is an interferon-inducible protein. Moreover, Ap Q A appeared to be a preferred substrate for 2-5A synthetase, also interferoninducible, priming the synthesis of 2P adenylated derivatives of Ap Q A, which in turn may serve as substrates of Fhit. Tumour suppressor activity of Fhit is assumed to be associated with involvement of the FhitWAp Q A complex in cytokine signalling pathway(s) controlling cell proliferation. The Ap n A family is potentially a novel class of signal-transducing molecules whose functions are yet to be determined.z 1998 Federation of European Biochemical Societies.