Instituto de Investigación Médica Mercedes y Martín Ferreyra, INIMEC - Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad Nacional de Córdoba, Córdoba, Argentina.
Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires (FCEyN-UBA) - Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales (IQUIBICEN) CONICET, Ciudad Autónoma de Buenos Aires, Argentina.
Acta Trop. 2024 Sep;257:107295. doi: 10.1016/j.actatropica.2024.107295. Epub 2024 Jun 19.
Histone post-translational modifications are extensively studied for their role in regulating gene transcription and cellular environmental adaptation. Research into these modifications has recently begun in the protozoan parasite Giardia lamblia, focusing on histone-modifying enzymes and specific post-translational changes. In the transformation from the trophozoite to the cyst form in the life cycle of this parasite, significant morphological and genetic alterations occur, culminating in the synthesis of cyst wall proteins responsible for forming the protective cyst wall. It has been previously demonstrated that histone deacetylation is required during encystation and that the enzyme lysine methyltransferase 1 is involved in the upregulation of encystation. Our study aims to extend the analysis to lysine methyltransferase 2 (GlKMT2) function. For this, two constructs were generated: one that downregulate the expression of GLKMT2 via antisense (glkmt2-as transgenic cells) and the other overexpressing GlKMT2 (glkmt2-ha transgenic cells). We found that the glktm2-as transgenic cells showed an arrest in progress at the late encystation stage. Consequently, the number of cysts produced was lower than that of the control cells. On the other hand, we found that the overexpression of GlKMT2 acts as a negative mutant of the enzyme. In this way, these glktm2-ha transgenic cells showed the same behavior during growth and encystation as glkmt2-as transgenic cells. This interplay between different enzymes acting during encystation reveals the complex process behind the differentiation of the parasite. Understanding how these enzymes play their role during the encystation of the parasite would allow the design of inhibitors to control the parasite.
组蛋白翻译后修饰因其在调节基因转录和细胞环境适应中的作用而被广泛研究。最近,在原生动物寄生虫蓝氏贾第鞭毛虫中,针对组蛋白修饰酶和特定的翻译后修饰,开始了相关研究。在寄生虫生命周期中,从滋养体向囊前期的转化过程中,会发生显著的形态和遗传改变,最终导致合成负责形成保护性囊壁的囊壁蛋白。先前已经证明,在囊前期形成过程中需要去乙酰化组蛋白,并且赖氨酸甲基转移酶 1 酶参与囊前期的上调。我们的研究旨在进一步分析赖氨酸甲基转移酶 2 (GlKMT2) 的功能。为此,我们构建了两种构建体:一种通过反义下调 GLKMT2 的表达 (glkmt2-as 转基因细胞),另一种过表达 GlKMT2 (glkmt2-ha 转基因细胞)。我们发现,glktm2-as 转基因细胞在晚期囊前期停滞不前。因此,产生的囊数量比对照细胞少。另一方面,我们发现 GlKMT2 的过表达表现为该酶的负突变体。这样,这些 glktm2-ha 转基因细胞在生长和囊前期的行为与 glkmt2-as 转基因细胞相同。这些在囊前期发挥作用的不同酶之间的相互作用揭示了寄生虫分化背后的复杂过程。了解这些酶在寄生虫囊前期形成过程中的作用方式,将有助于设计抑制剂来控制寄生虫。