Hsp70 and Larval Thermotolerance in Drosophila melanogaster: How Much is Enough and When is More Too Much?

Heat shock proteins (Hsps) and other molecular chaperones perform diverse cellular roles (e.g., inducible thermotolerance) whose functional consequences are concentration dependent. We manipulated Hsp70 concentration quantitatively in intact larvae of Drosophila melanogaster to examine its effect on survival, developmental time and tissue damage after heat shock. Larvae of an extra-copy strain, which has 22 hsp70 copies, produced Hsp70 more rapidly and to higher concentrations than larvae of a control strain, which has the wild-type 10 copies of the gene. Increasing the magnitude and duration of pretreatment increased Hsp70 concentrations, improved tolerance of more severe stress, and reduced delays in development. Pretreatment, however, did not protect against acute tissue damage. For larvae provided a brief or mild intensity pretreatment, faster expression of Hsp70 in the extra-copy strain improved survival to adult and reduced tissue damage 21 h after heat shock. Negative effects on survival ensued in extra-copy larvae pretreated most intensely, but their overexpression of Hsp70 did not increase tissue damage. Because rapid expression to yield a low Hsp70 concentration benefits larvae but overexpression harms them, natural selection may balance benefits and costs of high and low expression levels in natural populations

Natural and Genetic Engineering of the Heat-Shock Protein Hsp70 in Drosophila melanogaster: Consequences for Thermotolerance

Larvae of the fruit fly, Drosophila melanogaster, live within necrotic fruit, a challenging environment in which larvae can experience severe thermal stress. One response to thermal stress, the expression of heat-shock proteins (Hsps), has evolved distinctively in this species; the gene encoding Hsp70 has undergone extensive duplication and accounts for the bulk of Hsps that are expressed upon heat shock. Genetic engineering of hsp70 copy number is sufficient to affect thermotolerance at some (but not all) life stages. Increases in Hsp70, moreover, can protect intact larvae against thermal inactivation of the enzyme alcohol dehydrogenase and thermal inhibition of feeding. Deleterious consequences of high levels of Hsp70, however, may limit further evolutionary proliferation of hsp70 genes. These findings illustrate how the perspectives of integrative and comparative biology, if applied to even well-studied model organisms, can lead to novel findings

Experimental Manipulation of the Cost of Thermal Acclimation in Drosophila melanogaster

Acclimation to environmental change can impose costs on organisms. One potential cost is the energy and nutrients consumed by a physiological response, e.g. the resources required for expression of heat-shock proteins (Hsps). We examined the significance of this cost by genetic manipulation. We isolated four isofemale lines from a Drosophila melanogaster population previously transformed with a hsp70-kcZ fusion. Lines were similar in Hsp70 expression but differed in P-galactosidase expression upon heat shock, and replicates of each line were reared on a high quantity and low quantity medium. Multiple heat shock reduced survival in all lines, but did not increase developmental time. Variation in expression of β-galactosidase among lines, which differed more than 4-fold in response to heat treatment, was unrelated to the decreased survival. Thus the predicted effects of β-galactosidase expression on components of fitness were not evident. The superimposition of costs upon those normal for acclimation had no effect on mortality or developmental time, even when resources were especially limiting

Heritability of Expression of the 70kd Heat-shock Protein in Drosophila melanogaster and its Relevance to the Evolution of Thermotolerance

The principle inducible heat-shock protein of Drosophila melanogasrer, Hsp70, contributes to thermotolerance throughout the entire life cycle of the species but may also reduce fitness in some life stages. In principle, selection might maximize the benefits of Hsp70 expression relative to its costs by adjusting the magnitude of Hsp70 expression for each life-cycle stage independently. Therefore we examined whether the magnitude of Hsp70 ex pre sion varied during the life cycle and the relationship of this variation to several life-history traits. For 28 isofemale lines derived from a single natural population, estimates of heritable variation in Hsp70 expression ranged between 0.25 and 0.49, and the association among variation in first- and third-instar larvae and in adults correlated highly. Thus, Hsp70 expression is genetically coupled at these developmental stages. A line engineered with extra copies of the hsp70 gene produced more Hsp70 and survived heat shock much better than did a control strain . Among natural lines, Hsp70 expression was only weakly related to tolerance of heat shock and to larva-to-adult survival and developmental time at permissive temperatures. Additionally, lines with high adult survival developed slowly as larvae, which is a possible trade-off. These and other findings suggest that trade-offs may maintain quantitative variation both in heat shock protein expression and in life-history traits that associate with thermotolerance

The stable roommates problem with ties

We study the variant of the well-known stable roommates problem in which participants are permitted to express ties in their preference lists. In this setting, more than one definition of stability is possible. Here we consider two of these stability criteria, so-called super-stability and weak stability. We present a linear–time algorithm for finding a super-stable matching if one exists, given a stable roommates instance with ties. This contrasts with the known NP-hardness of the analogous problem under weak stability. We also extend our algorithm to cope with preference lists that are incomplete and/or partially ordered. On the other hand, for a given stable roommates instance with ties and incomplete lists, we show that the weakly stable matchings may be of different sizes and the problem of finding a maximum cardinality weakly stable matching is NP-hard, though approximable within a factor of 2

Changes in Thermotolerance and Hsp70 Expression with Domestication in Drosophila melanogaster

To examine how the duration of laboratory domestication may affect Drosophila stocks used in studies of thermotolerance, we measured expression of the inducible heat-shock protein Hsp70 and survival after heat shock in D. melanogaster strains recently collected from nature and maintained in laboratory culture for up to 50 or more generations. After an initial increase in both Hsp70 expression and thermotolerance immediately after transfer to laboratory medium, both traits remained fairly constant over time and variation among strains persisted through laboratory domestication. Furthermore, variation in heat tolerance and Hsp70 expression did not correlate with the length of time populations evolved in the laboratory. Therefore, while environmental variation likely contributed most to early shifts in strain tolerance and Hsp70 expression, other population parameters, for example genetic drift, inbreeding, and selection likely affected these traits little. As long as populations are maintained with large numbers of individuals, the culture of insects in the laboratory may have little effect on the tolerance of different strains to thermal stress