McKechnie S W, Halford M M, McColl G, Hoffmann A A
Department of Genetics and Developmental Biology, Monash University, Victoria 3168, Australia.
Proc Natl Acad Sci U S A. 1998 Mar 3;95(5):2423-8. doi: 10.1073/pnas.95.5.2423.
Inducible heat shock genes are considered a major component of the molecular mechanisms that confer cellular protection against a variety of environmental stresses, in particular high temperature extremes. We have tested the association between expression of the heat shock RNA gene hsr-omega and thermoresistance by generating thermoresistant lines of Drosophila melanogaster after application of two distinct regimes of laboratory selection. One set of lines was selected for resistance to knockdown by heat stress and the other was similarly selected but before selection a mild heat exposure known to increase resistance (heat hardening) was applied. A cross between resistant and susceptible lines confirmed our earlier observation that increased thermal tolerance cosegregates with allelic variation in the hsr-omega gene. This cosegregating variation is attributed largely to two haplotype groups. Using quantitative reverse transcription-PCR, we find evidence for divergent phenotypic responses in the two selection regimes, involving both structural and regulatory changes in hsr-omega. Lines selected after hardening showed increased levels of the cytoplasmic transcript but decreased levels of the nuclear transcript. Lines selected without hardening showed decreased levels of the cytoplasmic transcript. The allelic frequency changes at hsr-omega could not by themselves account for the altered transcription patterns. Our results support the idea that the functional RNA molecules transcribed from hsr-omega are an important and polymorphic regulatory component of an insect thermoresistance phenotype.
可诱导热休克基因被认为是赋予细胞抵御多种环境压力(尤其是极端高温)保护作用的分子机制的主要组成部分。我们通过应用两种不同的实验室选择方案培育出抗热的黑腹果蝇品系,测试了热休克RNA基因hsr-omega的表达与耐热性之间的关联。一组品系被选择用于抵抗热应激导致的击倒,另一组品系也进行了类似选择,但在选择之前进行了已知能增加抗性的轻度热暴露(热硬化)。抗性品系和敏感品系之间的杂交证实了我们早期的观察结果,即热耐受性的增加与hsr-omega基因的等位基因变异共分离。这种共分离变异主要归因于两个单倍型组。使用定量逆转录PCR,我们发现了两种选择方案中不同表型反应的证据,涉及hsr-omega的结构和调控变化。硬化后选择的品系显示细胞质转录本水平增加,但核转录本水平降低。未进行硬化选择的品系显示细胞质转录本水平降低。hsr-omega处的等位基因频率变化本身无法解释转录模式的改变。我们的结果支持这样一种观点,即从hsr-omega转录的功能性RNA分子是昆虫耐热性表型的一个重要且多态的调控成分。