Onorato L, Stirmer B, Showe M K
J Virol. 1978 Aug;27(2):409-26. doi: 10.1128/JVI.27.2.409-426.1978.
To determine the function of individual gene products in the assembly and maturation of the T4 prehead, we have isolated and characterized aberrant preheads produced by mutations in three of the T4 head genes. Mutants in gene 21, which codes for the T4 maturation proteases, produce rather stable preheads whose morphology and protein composition are consistent with a wild-type prehead blocked in the maturation cleavages. Mutants in gene 24 produce similar structures which are unstable because they have gaps at all of their icosahedral vertices except the membrane attachment site. In addition, greatly elongated "giant preheads" are produced, suggesting that in the absence of P24 at the vertices, the distal cap of the prehead is unstable, allowing abnormal elongation of broth the prehead core and its shell. Vertex completion by P24 is required to allow the maturation cleavages to occur, and 24- preheads can be matured to capsids in vitro by the addition of P24. Preheads produced by a temperature-sensitive mutant in gene 23 are deficient in core proteins. We show that the shell of these preheads has the expanded lattice characteristic of the mature capsid as well as the binding sites for the proteins hoc and soc, even though none of the maturation cleavage takes place. We also show that 21- preheads composed of wild-type P23 can be expanded in vitro without cleavage.
为了确定单个基因产物在T4前头部组装和成熟过程中的功能,我们分离并鉴定了由T4头部三个基因的突变产生的异常前头部。编码T4成熟蛋白酶的基因21中的突变体产生相当稳定的前头部,其形态和蛋白质组成与在成熟切割中受阻的野生型前头部一致。基因24中的突变体产生类似的结构,这些结构不稳定,因为除了膜附着位点外,它们在所有二十面体顶点处都有间隙。此外,还产生了大大拉长的“巨型前头部”,这表明在顶点处没有P24的情况下,前头部的远端帽不稳定,从而允许前头部核心及其外壳异常伸长。P24完成顶点封闭是使成熟切割发生所必需的,并且通过添加P24,24-前头部可以在体外成熟为衣壳。基因23中的温度敏感突变体产生的前头部缺乏核心蛋白。我们表明,这些前头部的外壳具有成熟衣壳的扩展晶格特征以及hoc和soc蛋白的结合位点,尽管没有发生任何成熟切割。我们还表明,由野生型P23组成的21-前头部可以在体外无切割地扩展。