Bartholomeu Daniella C, Cerqueira Gustavo C, Leão Ana Carolina A, daRocha Wanderson D, Pais Fabiano S, Macedo Camila, Djikeng Appolinaire, Teixeira Santuza M R, El-Sayed Najib M
Department of Parasitology, Federal University of Minas Gerais, Belo Horizonte, MG, Brazil.
Nucleic Acids Res. 2009 Jun;37(10):3407-17. doi: 10.1093/nar/gkp172. Epub 2009 Mar 31.
A novel large multigene family was recently identified in the human pathogen Trypanosoma cruzi, causative agent of Chagas disease, and corresponds to approximately 6% of the parasite diploid genome. The predicted gene products, mucin-associated surface proteins (MASPs), are characterized by highly conserved N- and C-terminal domains and a strikingly variable and repetitive central region. We report here an analysis of the genomic organization and expression profile of masp genes. Masps are not randomly distributed throughout the genome but instead are clustered with genes encoding mucin and other surface protein families. Masp transcripts vary in size, are preferentially expressed during the trypomastigote stage and contain highly conserved 5' and 3' untranslated regions. A sequence analysis of a trypomastigote cDNA library reveals the expression of multiple masp variants with a bias towards a particular masp subgroup. Immunofluorescence assays using antibodies generated against a MASP peptide reveals that the expression of particular MASPs at the cell membrane is limited to subsets of the parasite population. Western blots of phosphatidylinositol-specific phospholipase C (PI-PLC)-treated parasites suggest that MASP may be GPI-anchored and shed into the medium culture, thus contributing to the large repertoire of parasite polypeptides that are exposed to the host immune system.
最近在人类病原体克氏锥虫(恰加斯病的病原体)中发现了一个新的大型多基因家族,约占该寄生虫二倍体基因组的6%。预测的基因产物,即粘蛋白相关表面蛋白(MASP),其特征是具有高度保守的N端和C端结构域以及一个显著可变且重复的中央区域。我们在此报告对masp基因的基因组组织和表达谱的分析。Masp并非随机分布于整个基因组,而是与编码粘蛋白和其他表面蛋白家族的基因聚集在一起。Masp转录本大小各异,在锥鞭毛体阶段优先表达,并且包含高度保守的5'和3'非翻译区。对锥鞭毛体cDNA文库的序列分析揭示了多个masp变体的表达,且偏向于特定的masp亚组。使用针对MASP肽产生的抗体进行的免疫荧光测定表明,特定MASP在细胞膜上的表达仅限于寄生虫群体的亚群。对经磷脂酰肌醇特异性磷脂酶C(PI-PLC)处理的寄生虫进行的蛋白质免疫印迹分析表明,MASP可能通过糖基磷脂酰肌醇(GPI)锚定并释放到培养基中,从而增加了暴露于宿主免疫系统的寄生虫多肽种类。