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Eukaryotic GPN-loop GTPases paralogs use a dimeric assembly reminiscent of archeal GPN

机译:真核GPN环GTPases旁系同源物使用的二聚体让人想起古细菌GPN

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摘要

GTP ases are molecular switches that regulate a wide-range of cellular processes. The GPN-loop GTP ase (GPN) is a sub-family of P-loop NTP ase that evolved from a single gene copy in archaea to triplicate paralog genes in eukaryotes, each having a non-redundant essential function in cell. In Saccharomyces cerevisiae, yGPN1 and yGPN2 are involved in sister chromatid cohesion mechanism, whereas nothing is known regarding yGPN3 function. Previous high-throughput experiments suggested that GPN paralogs interaction may occur. In this work, GPN|GPN contact was analyzed in details using TAPTag approach, yeast two-hybrid assay, in silico energy computation and site-directed mutagenesis of a conserved Glu residue located at the center of the interaction interface. It is demonstrated that this residue is essential for cell viability. A chromatid cohesion assay revealed that, like yGPN1 and yGPN2, yGPN3 also plays a role in sister chromatid cohesion. These results suggest that all three GPN proteins act at the molecular level in sister chromatid cohesion mechanism as a GPN|GPN complex reminiscent of the homodimeric structure of PAB0955, an archaeal member of GPN-loop GTP ase. ? 2013 Landes Bioscience.
机译:GTP驴是调节多种细胞过程的分子开关。 GPN环GTP酶(GPN)是P环NTP酶的一个亚家族,它从古细菌中的单个基因拷贝演变成真核生物中的平行同源基因一式三份,每个基因在细胞中均具有非冗余的基本功能。在酿酒酵母中,yGPN1和yGPN2参与了姐妹染色单体的内聚机制,而关于yGPN3的功能则一无所知。先前的高通量实验表明,可能发生GPN旁系同源物相互作用。在这项工作中,使用TAPTag方法,酵母双杂交测定,计算机能量计算和位于相互作用界面中心的保守Glu残基的定向诱变,详细分析了GPN | GPN接触。证明该残基对于细胞生存力是必不可少的。染色单体内聚测定表明,与yGPN1和yGPN2一样,yGPN3在姐妹染色单体内聚中也起作用。这些结果表明,所有三种GPN蛋白在姐妹染色单体内聚机制中均在分子水平上作为GPN | GPN复合物起作用,使人联想到GPN环GTP酶的古细菌成员PAB0955的同二聚体结构。 ? 2013 Landes生物科学。

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