Afp14 is involved in regulating the length of Anti-feeding prophage (Afp)
Afp14 is involved in regulating the length of Anti-feeding prophage (Afp)
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DOI：
10.1111/mmi.12974
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发表时间：
2015-05-01
影响因子：
3.6
通讯作者：
Hurst, Mark R. H.
中科院分区：
文献类型：
--
作者：
Rybakova, Daria;Schramm, Peter;Hurst, Mark R. H.
抗摄食原噬菌体（AFP）是一种噬菌体尾样颗粒，可引起新西兰草螟Costelytra zealandica的摄食停止，由18个开放阅读框架（AFP 1 -18）编码。afp 14的C-末端截短导致AFP颗粒缩短，这表明Afp 14参与了AFP长度的确定。我们构建了一个Afp组装系统（afp 1 -18），其中Afp 14在N-末端88个残基后被截短。当该构建体在表达N-末端98-氨基酸Afp 14构建体的大肠杆菌中反式表达时，产生完全组装的Afp，但是在N-末端96-氨基酸Afp 14构建体的情况下没有检测到组装的Afp。这些结果表明，98 N-末端，氨基酸残基的Afp 14是至关重要的启动Afp组装通过基板形成。野生型afp 14的基于反式的表达导致不同长度的AFP颗粒，所有这些都比野生型AFP颗粒短。另一方面，类似的表达的AFP 14窝藏C-末端延伸（KLLEH 6）导致延长的AFP颗粒。这些信息，结合生物信息学数据，使我们能够提出一个模型描绘的机制和作用的Afp 14在成熟的AFP颗粒。
The anti-feeding prophage (Afp), a phage-tail-like particle that causes cessation of feeding in the New Zealand grass grub, Costelytra zealandica, is encoded by 18 open reading frames (afp1-18). C-terminal truncations of afp14 resulted in shortened Afp particles, suggesting that Afp14 is involved in Afp length determination. We constructed an Afp assembly system (afp1-18), wherein Afp14 was truncated after the N-terminal 88 residues. This construct, when expressed in trans in Escherichia coli expressing a N-terminal 98-amino acid Afp14 construct, yielded fully assembled Afp but no assembled Afp was detected in the case of a N-terminal 96-amino acid Afp14 construct. These results suggested that the 98 N-terminal, amino acid residues of Afp14 is crucial for the initiation of Afp assembly via baseplate formation. Trans-based expression of wild-type afp14 resulted in Afp particles of varying lengths, all of which were shorter than the wild-type Afp particle. On the other hand, similar expression of Afp14 harboring a C-terminal extension (KLLEH6) resulted in elongated Afp particles. This information, combined with bioinformatics data, allowed us to propose a model delineating the mechanism and role of Afp14 in the maturation of the Afp particle.