Theodoraki M A, Mintzas A C
Division of Genetics, Cell and Developmental Biology, Department of Biology, University of Patras, Patras, Greece.
Insect Mol Biol. 2006 Dec;15(6):839-52. doi: 10.1111/j.1365-2583.2006.00691.x.
This report presents the cDNA cloning, heat shock regulation and developmental expression of the hsp90 gene homologue of the Mediterranean fruit fly Ceratitis capitata (medfly). The isolated cDNA contained the coding region, the 3'UTR and most of the 5'UTR of the medfly hsp90 homologue, which was named Cchsp83. The deduced CcHSP83 polypeptide contained all the highly conserved amino acid segments that characterize the cytosolic members of the HSP90 family. Genomic analysis showed that the Cchsp83 gene is unique and was mapped at the 94C division of the sixth polytene chromosome. The size of the Cchsp83 mRNA was found to be approximately 2.7 kb. The predicted molecular mass of the CcHSP83 protein was 81.4 kDa, while the apparent molecular weight estimated by SDS-PAGE was approximately 90 kDa. Phylogenetic analysis based on 14 insect HSP90 amino acid sequences was consistent with the known phylogeny at low taxonomic level. The Cchsp83 gene is constitutively expressed in all stages of medfly development and is induced from a low level to several-fold by heat, depending on the developmental stage. Heat shock induction begins at 30 degrees C, reaching a maximum between 35 and 41 degrees C. Cchsp83 RNA expression is highly regulated during embryonic development; however, the temporal fluctuations in RNA levels during embryogenesis were not followed by similar fluctuations in the levels of the protein.
本报告介绍了地中海实蝇（Ceratitis capitata，medfly）hsp90基因同源物的cDNA克隆、热休克调节及发育表达情况。分离得到的cDNA包含地中海实蝇hsp90同源物的编码区、3'非翻译区（UTR）以及大部分5'UTR，该同源物被命名为Cchsp83。推导的CcHSP83多肽包含了所有高度保守的氨基酸片段，这些片段是HSP90家族胞质成员的特征。基因组分析表明，Cchsp83基因是唯一的，定位于第六条多线染色体的94C区。发现Cchsp83 mRNA的大小约为2.7 kb。预测的CcHSP83蛋白分子量为81.4 kDa，而通过SDS-PAGE估计的表观分子量约为90 kDa。基于14种昆虫HSP90氨基酸序列的系统发育分析与低分类水平下已知的系统发育一致。Cchsp83基因在medfly发育的所有阶段均组成性表达，并根据发育阶段受热诱导，从低水平诱导至几倍。热休克诱导在30℃开始，在35至41℃之间达到最大值。Cchsp83 RNA表达在胚胎发育过程中受到高度调节；然而，胚胎发生过程中RNA水平的时间波动并未伴随着蛋白质水平的类似波动。
