Lal Shardul Vikram, Brahma Biswajit, Gohain Moloya, Mohanta Debashish, De Bidan Chandra, Chopra Meenu, Dass Gulshan, Vats Ashutosh, Upadhyay Ramesh C, Datta T K, De Sachinandan
Animal Genomics Lab, Animal Biotechnology Centre, National Dairy Research Institute, Karnal, 132001, Haryana, India,
Cell Stress Chaperones. 2015 May;20(3):545-54. doi: 10.1007/s12192-014-0563-y. Epub 2015 Feb 6.
In eukaryotes, the heat shock factors (HSFs) are recognized as the master regulator of the heat shock response. In this respect, the genes encoding the heat shock factors seem to be important for adaptation to thermal stress in organisms. Despite this, only few mammalian HSFs has been characterized. In this study, four major heat shock factor genes viz. HSF-1, 2, 4, and 5 were studied. The main objective of the present study was to characterize the cDNA encoding using conserved gene specific primers and to investigate the expression status of these buffalo HSF genes. Our RT-PCR analysis uncovered two distinct variants of buffalo HSF-1 and HSF-2 gene transcripts. In addition, we identified a variant of the HSF5 transcript in buffalo lacking a DNA-binding domain. In silico analysis of deduced amino acid sequences for buffalo HSF genes showed domain architecture similar to other mammalian species. Changes in the gene expression profile were noted by quantitative real-time PCR (qRT-PCR) analysis. We detected the transcript of buffalo HSF genes in different tissues. We also evaluated the seasonal changes in the expression of HSF genes. Interestingly, the transcript level of HSF-1 gene was found upregulated in months of high and low ambient temperatures. In contrast, the expression of the HSF-4 and 5 genes was found to be downregulated in months of high ambient temperature. This suggests that the intricate balance of different HSFs is adjusted to minimize the effect of seasonal changes in environmental conditions. These findings advance our understanding of the complex, context-dependent regulation of HSF gene expression under normal and stressful conditions.
在真核生物中，热休克因子（HSFs）被认为是热休克反应的主要调节因子。在这方面，编码热休克因子的基因似乎对生物体适应热应激很重要。尽管如此，只有少数哺乳动物的热休克因子得到了表征。在本研究中，对四个主要的热休克因子基因，即HSF-1、2、4和5进行了研究。本研究的主要目的是使用保守的基因特异性引物对编码的cDNA进行表征，并研究这些水牛热休克因子基因的表达状态。我们的逆转录聚合酶链反应（RT-PCR）分析发现了水牛HSF-1和HSF-2基因转录本的两种不同变体。此外，我们在水牛中鉴定出一种缺少DNA结合结构域的HSF5转录本变体。对水牛热休克因子基因推导的氨基酸序列进行的电子分析显示，其结构域结构与其他哺乳动物物种相似。通过定量实时聚合酶链反应（qRT-PCR）分析注意到基因表达谱的变化。我们在不同组织中检测到了水牛热休克因子基因的转录本。我们还评估了热休克因子基因表达的季节性变化。有趣的是，发现HSF-1基因的转录水平在环境温度高和低的月份上调。相反，发现HSF-4和5基因的表达在环境温度高的月份下调。这表明不同热休克因子的复杂平衡被调整，以尽量减少环境条件季节性变化的影响。这些发现推进了我们对正常和应激条件下热休克因子基因表达的复杂、依赖背景的调控的理解。
