Pugliese Luciana, Georg Raphaela C, Fietto Luciano G, Gomes Suely L
Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, Brazil.
Gene. 2008 Mar 31;411(1-2):59-68. doi: 10.1016/j.gene.2008.01.005. Epub 2008 Jan 24.
HSP90 proteins are important molecular chaperones involved in multiple cellular processes. This work reports the characterization of cDNAs encoding two distinct HSP90 proteins (named HSP90A and HSP90B) from the chytridiomycete Blastocladiella emersonii. Deduced amino acid sequences of HSP90A and HSP90B exhibit signatures of the cytosolic and endoplasmic reticulum (ER) HSP90 proteins, respectively. A genomic clone encoding HSP90A was also characterized indicating the presence of a single intron of 184 bp interrupting the coding region, located near the amino-terminus of the protein. Expression of both HSP90A and HSP90B genes increases significantly during heat shock at 38 degrees C, with highest induction ratios observed in cells stressed during germination of the fungus. Changes in the amount of HSP90A transcript were also evaluated during B. emersonii life cycle at physiological temperature (27 degrees C), and its levels were found to increase both during germination and sporulation of the fungus. HSP90A protein levels were analyzed during B. emersonii life cycle and significant changes were observed only during sporulation. Furthermore, during heat stress a large increase in the amount of HSP90A protein was observed. Induction of HSP90A and HSP90B genes during heat stress indicates the importance of both genes in the response to high temperature in B. emersonii.
热休克蛋白90(HSP90)是参与多种细胞过程的重要分子伴侣。这项工作报道了来自壶菌埃默森芽枝霉中编码两种不同HSP90蛋白(命名为HSP90A和HSP90B)的cDNA的特征。HSP90A和HSP90B推导的氨基酸序列分别显示出胞质和内质网(ER)HSP90蛋白的特征。一个编码HSP90A的基因组克隆也被鉴定,表明在靠近蛋白质氨基末端的编码区存在一个184bp的单一内含子。在38℃热休克期间,HSP90A和HSP90B基因的表达均显著增加,在真菌萌发期间受到胁迫的细胞中观察到最高的诱导率。在埃默森芽枝霉生命周期的生理温度(27℃)下,也评估了HSP90A转录本数量的变化,发现其水平在真菌萌发和孢子形成期间均增加。在埃默森芽枝霉生命周期中分析了HSP90A蛋白水平,仅在孢子形成期间观察到显著变化。此外,在热胁迫期间,观察到HSP90A蛋白的量大幅增加。热胁迫期间HSP90A和HSP90B基因的诱导表明这两个基因在埃默森芽枝霉对高温的反应中都很重要。