New Insights into the Structure and Assembly of Bacteriophage P1.
New Insights into the Structure and Assembly of Bacteriophage P1.
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DOI：
10.3390/v14040678
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发表时间：
2022-03-25
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影响因子：
--
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噬菌体P1是大肠杆菌的主要转导噬菌体。杆菌尽管它在推进E.由于P1基因的遗传学研究尚不成熟，现代分子生物学技术还没有应用于P1基因的结构研究。在这里，我们报告的蛋白质组的P1病毒体，通过液相色谱串联质谱法测定。此外，单基因敲除文库鉴定了以下五个先前未知的必需基因：pmgA、pmgB、pmgC、pmgG和pmgR。此外，主要衣壳蛋白的蛋白水解加工是P1形态发生的已知特征，我们通过N-末端测序确定了E120和S121之间的加工位点，产生448个残基，49.3 kDa的成熟肽。此外，P1防御限制（Dar）系统由六种已知的蛋白质组成，这些蛋白质在形态发生过程中被掺入病毒体中。其中最大的是DarB，是一种250 kDa的蛋白质，据信在感染期间转移到细胞中。DarB缺失表明存在N-末端包装信号，并且显示DarB的N-末端30个残基足以将异源报告蛋白引导至衣壳。总的来说，这些数据扩展了基本的结构P1蛋白，并将P1作为细胞递送的纳米机器引入。
Bacteriophage P1 is the premier transducing phage of E. coli. Despite its prominence in advancing E. coli genetics, modern molecular techniques have not been applied to thoroughly understand P1 structure. Here, we report the proteome of the P1 virion as determined by liquid chromatography tandem mass-spectrometry. Additionally, a library of single-gene knockouts identified the following five previously unknown essential genes: pmgA, pmgB, pmgC, pmgG, and pmgR. In addition, proteolytic processing of the major capsid protein is a known feature of P1 morphogenesis, and we identified the processing site by N-terminal sequencing to be between E120 and S121, producing a 448-residue, 49.3 kDa mature peptide. Furthermore, the P1 defense against restriction (Dar) system consists of six known proteins that are incorporated into the virion during morphogenesis. The largest of these, DarB, is a 250 kDa protein that is believed to translocate into the cell during infection. DarB deletions indicated the presence of an N-terminal packaging signal, and the N-terminal 30 residues of DarB are shown to be sufficient for directing a heterologous reporter protein to the capsid. Taken together, the data expand on essential structural P1 proteins as well as introduces P1 as a nanomachine for cellular delivery.
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影响因子：
3.7
作者：
Liu J;Chen CY;Shiomi D;Niki H;Margolin W
通讯作者：
Margolin W
影响因子：
3.7
作者：
IIDA, S;STREIFF, MB;ARBER, W
通讯作者：
ARBER, W
影响因子：
4.8
作者：
Boulanger, Pascale;Jacquot, Pierre;Letellier, Lucienne
通讯作者：
Letellier, Lucienne
影响因子：
5.6
作者：
ARBER, W;DUSSOIX, D
通讯作者：
DUSSOIX, D
影响因子：
5.6
作者：
Effantin, G.;Boulanger, P.;Conway, J. F.
通讯作者：
Conway, J. F.