The Small Ubiquitin-Like Modifier (SUMO) and SUMO-Conjugating System of Chlamydomonas reinhardtii
The Small Ubiquitin-Like Modifier (SUMO) and SUMO-Conjugating System of Chlamydomonas reinhardtii
复制标题
DOI：
10.1534/genetics.108.089128
复制
发表时间：
2008-05
期刊：
影响因子：
3.3
通讯作者：
Y. Wang;I. Ladunga;Amy R. Miller;K. Horken;Thomas M. Plucinak;D. Weeks;C. Bailey
中科院分区：
文献类型：
--
作者：
Y. Wang;I. Ladunga;Amy R. Miller;K. Horken;Thomas M. Plucinak;D. Weeks;C. Bailey
莱茵衣藻基因组的完整DNA序列和先进的计算生物学工具的可用性，允许阐明和研究的小泛素样修饰剂（SUMO）系统在这个单细胞光合细菌和模型真核细胞系统。SUMO是泛素样蛋白超家族的成员，其作为翻译后修饰共价连接至靶蛋白，以响应细胞环境的变化而改变靶蛋白的定位、稳定性和/或功能。通过不同的基因预测、隐马尔可夫模型和数据库搜索工具从智人、酿酒酵母和拟南芥中发现了三个SUMO同源物（CrSUMO 96、CrSUMO 97和CrSUMO 148）和三个新的SUMO相关蛋白（CrSUMO-like 89 A、CrSUMO-like 89 B和CrSUMO-like 90）。其中CrSUMO 96为A.拟南芥抗SUMO 1抗体。通过免疫沉淀法纯化游离CrSUMO 96，并通过质谱分析进行鉴定。在C.研究表明，莱茵衣藻在基于大肠杆菌的体内嵌合SUMO化系统中具有功能。CrSUMO 96抗体在全细胞提取物和核定位SUMO化蛋白质的Western印迹分析中识别游离和缀合形式的CrSUMO 96，具有原位免疫荧光。Western blot分析表明，当细胞受到环境胁迫，如热休克和渗透胁迫时，SUMO结合蛋白显着增加。相关分析显示，C.莱茵衣藻基因组
The availability of the complete DNA sequence of the Chlamydomonas reinhardtii genome and advanced computational biology tools has allowed elucidation and study of the small ubiquitin-like modifier (SUMO) system in this unicellular photosynthetic alga and model eukaryotic cell system. SUMO is a member of a ubiquitin-like protein superfamily that is covalently attached to target proteins as a post-translational modification to alter the localization, stability, and/or function of the target protein in response to changes in the cellular environment. Three SUMO homologs (CrSUMO96, CrSUMO97, and CrSUMO148) and three novel SUMO-related proteins (CrSUMO-like89A, CrSUMO-like89B, and CrSUMO-like90) were found by diverse gene predictions, hidden Markov models, and database search tools inferring from Homo sapiens, Saccharomyces cerevisiae, and Arabidopsis thaliana SUMOs. Among them, CrSUMO96, which can be recognized by the A. thaliana anti-SUMO1 antibody, was studied in detail. Free CrSUMO96 was purified by immunoprecipitation and identified by mass spectrometry analysis. A SUMO-conjugating enzyme (SCE) (E2, Ubc9) in C. reinhardtii was shown to be functional in an Escherichia coli-based in vivo chimeric SUMOylation system. Antibodies to CrSUMO96 recognized free and conjugated forms of CrSUMO96 in Western blot analysis of whole-cell extracts and nuclear localized SUMOylated proteins with in situ immunofluorescence. Western blot analysis showed a marked increase in SUMO conjugated proteins when the cells were subjected to environmental stresses, such as heat shock and osmotic stress. Related analyses revealed multiple potential ubiquitin genes along with two Rub1 genes and one Ufm1 gene in the C. reinhardtii genome.