Prohead RNA of bacteriophage phi 29: size, stoichiometry and biological activity.
Prohead RNA of bacteriophage phi 29: size, stoichiometry and biological activity.
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噬菌体 phi 29 的前体 RNA：大小、化学计量和生物活性。
DOI：
10.1093/nar/17.9.3459
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发表时间：
1989
影响因子：
14.9
通讯作者：
Anderson,D
中科院分区：
文献类型：
--
作者：
Wichitwechkarn,J;Bailey,S;Bodley,JW;Anderson,D
我们先前证明了（Guoet al.，1987. Nucl. Acids Res.15，7081-7090），纯化的噬菌体π29的前头部含有DNA包装所必需的120个碱基的RNA。在这里，我们报告说，这种RNA存在主要是作为一个聚合物的钙。174个残基的噬菌体感染的细胞和thatca。在纯化前体的过程中，外源核酸酶从其3′-末端切割54个碱基。在体外DNA包装和噬菌体组装中，长、短RNA的活性相似。我们报告了RNA长型的序列，并表明可以从π29的亲戚φ21，φ15和SF 5的前体中分离出类似的长型和短型。无RNA的前头部的重建中的浓度依赖性表明，一个拷贝的RNA足以恢复DNA包装活性的无RNA的前头部。然而，定量测量表明，5至6个拷贝的RNA存在于从噬菌体感染的细胞分离的前体上。
We previously demonstrated (Guoet al., 1987. Nucl. Acids Res. 15, 7081–7090) that purified proheads of bacteriophageπ29 contain an RNA of 120 bases which is essential for DNA packaging. Here we report that this RNA exists primarily as a polymer ofca. 174 residues in phage-infected cells and thatca. 54 bases are cleaved from its 3′-terminus by adventitious nucleases during the purification of proheads. The long and short forms of the RNA had similar activity inin vitroDNA packaging and phage assembly. We report the sequence of the long form of the RNA and show that similar long and short forms can be isolated from the proheads of theπ29 relativesφ21,φ15 and SF5. The concentration dependence in the reconstitution of RNA-free proheads suggests that one copy of the RNA is sufficient to restore DNA packaging activity to RNA-free proheads. However, quantitative measurements indicate that 5 to 6 copies of the RNA are present on proheads isolated from phage-infected cells.