The GTP-binding protein YlqF participates in the late step of 50 S ribosomal subunit assembly in Bacillus subtilis
The GTP-binding protein YlqF participates in the late step of 50 S ribosomal subunit assembly in Bacillus subtilis
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DOI：
10.1074/jbc.m512556200
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发表时间：
2006-03-24
影响因子：
4.8
通讯作者：
Ogasawara, N
中科院分区：
文献类型：
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作者：
Matsuo, Y;Morimoto, T;Ogasawara, N
枯草芽孢杆菌YlqF属于Era/Obg亚家族的小分子GTP结合蛋白，是细菌生长所必需的。在此，我们报道了YlqF参与了50个S核糖体亚基组装的后期。YlqF与50个S亚基共分，这取决于不可切割的GTP类似物的存在。体外实验表明，50个S亚基对YlqF的GTP酶活性有明显的刺激作用。硫酸二甲酯足迹分析表明，YlqF结合在23个S rRNA中的一个独特位置。酵母双杂交数据显示YlqF与枯草杆菌L25蛋白(CTC)之间存在相互作用。通过对纯化的蛋白质进行下拉实验证实了这种相互作用。具体来说，YlqF位于S 50亚基上的A位和P位附近。对YlqF缺失细胞中聚集的异常的50个S亚基的蛋白质组分析表明，位于YlqF结合域附近的L16和L27蛋白缺失。我们的结果综合表明，YlqF将组织50个S核糖体亚基组装的后期。
Bacillus subtilis YlqF belongs to the Era/Obg subfamily of small GTP-binding proteins and is essential for bacterial growth. Here we report that YlqF participates in the late step of 50 S ribosomal subunit assembly. YlqF was co-fractionated with the 50 S subunit, depending on the presence of noncleavable GTP analog. Moreover, the GTPase activity of YlqF was stimulated specifically by the 50 S subunit in vitro. Dimethyl sulfate footprinting analysis disclosed that YlqF binds to a unique position in 23 S rRNA. Yeast two-hybrid data revealed interactions between YlqF and the B. subtilis L25 protein (Ctc). The interaction was confirmed by the pull-down assay of the purified proteins. Specifically, YlqF is positioned around the A-site and P-site on the 50 S subunit. Proteome analysis of the abnormal 50 S subunits that accumulated in YlqF-depleted cells showed that L16 and L27 proteins, located near the YlqF-binding domain, are missing. Our results collectively indicate that YlqF will organize the late step of 50 S ribosomal subunit assembly.