Kolakowski Damian, Rzepnikowska Weronika, Kaniak-Golik Aneta, Zoladek Teresa, Kaminska Joanna
Institute of Biochemistry and Biophysics, Polish Academy of Science, 02-106 Warsaw, Poland.
Neuromuscular Unit, Mossakowski Medical Research Institute, Polish Academy of Sciences, 02-106 Warsaw, Poland.
Int J Mol Sci. 2021 Nov 12;22(22):12274. doi: 10.3390/ijms222212274.
VPS13 proteins are evolutionarily conserved. Mutations in the four human genes () encoding VPS13A-D proteins are linked to developmental or neurodegenerative diseases. The relationship between the specific localization of individual VPS13 proteins, their molecular functions, and the pathology of these diseases is unknown. Here we used a yeast model to establish the determinants of Vps13's interaction with the membranes of Golgi apparatus. We analyzed the different phenotypes of the Δ ∆ strain, with reduced activity of the Arf1 GTPase, the master regulator of Golgi function and entirely devoid of Vps13. Our analysis led us to propose that Vps13 and Arf1 proteins cooperate at the Golgi apparatus. We showed that Vps13 binds to the Arf1 GTPase through its C-terminal Pleckstrin homology (PH)-like domain. This domain also interacts with phosphoinositol 4,5-bisphosphate as it was bound to liposomes enriched with this lipid. The homologous domain of VPS13A exhibited the same behavior. Furthermore, a fusion of the PH-like domain of Vps13 to green fluorescent protein was localized to Golgi structures in an Arf1-dependent manner. These results suggest that the PH-like domains and Arf1 are determinants of the localization of VPS13 proteins to the Golgi apparatus in yeast and humans.
VPS13蛋白在进化上是保守的。编码VPS13A - D蛋白的四个人类基因（）中的突变与发育性或神经退行性疾病有关。单个VPS13蛋白的特定定位、它们的分子功能与这些疾病病理学之间的关系尚不清楚。在这里，我们使用酵母模型来确定Vps13与高尔基体膜相互作用的决定因素。我们分析了Arf1 GTPase活性降低且完全缺乏Vps13的ΔΔ菌株的不同表型。我们的分析使我们提出Vps13和Arf1蛋白在高尔基体处协同作用。我们表明Vps13通过其C末端类普列克底物蛋白同源（PH）结构域与Arf1 GTPase结合。该结构域还与磷脂酰肌醇4,5 - 二磷酸相互作用，因为它与富含这种脂质的脂质体结合。VPS13A的同源结构域表现出相同的行为。此外，Vps13的类PH结构域与绿色荧光蛋白的融合体以Arf1依赖的方式定位于高尔基体结构。这些结果表明，类PH结构域和Arf1是酵母和人类中VPS13蛋白定位于高尔基体的决定因素。
