Anderson L M, Bott K F
J Virol. 1985 Jun;54(3):773-80. doi: 10.1128/JVI.54.3.773-780.1985.
Defective bacteriophage PBSX, a resident of all Bacillus subtilis 168 chromosomes, packages fragments of DNA from all portions of the host chromosome when induced by mitomycin C. In this study, the physical process for DNA packaging of both chromosomal and plasmid DNAs was examined. Discrete 13-kilobase (kb) lengths of DNA were packaged by wild-type phage, and the process was DNase I resistant and probably occurred by a head-filling mechanism. Genetically engineered isogenic host strains having a chloramphenicol resistance determinant integrated as a genetic flag at two different regions of the chromosome were used to monitor the packaging of specific chromosomal regions. No dramatic selectivity for these regions could be documented. If the wild-type strain 168 contains autonomously replicating plasmids, especially pC194, the mitomycin C induces an increase in size of resident plasmid DNA, which is then packaged as 13-kb pieces into phage heads. In strain RB1144, which lacks substantial portions of the PBSX resident phage region, mitomycin C treatment did not affect the structure of resident plasmids. Induction of PBSX started rolling circle replication on plasmids, which then became packaged as 13-kb fragments. This alteration or cannibalization of plasmid replication resulting from mitomycin C treatment requires for its function some DNA within the prophage deletion of strain RB1144.
缺陷噬菌体PBSX是所有枯草芽孢杆菌168染色体中的一种常驻噬菌体,当用丝裂霉素C诱导时,它会包装来自宿主染色体所有部分的DNA片段。在本研究中,对染色体DNA和质粒DNA的包装物理过程进行了研究。野生型噬菌体包装的DNA长度为离散的13千碱基(kb),该过程对DNase I具有抗性,可能是通过头部填充机制发生的。使用在染色体的两个不同区域整合了氯霉素抗性决定簇作为遗传标记的基因工程同基因宿主菌株来监测特定染色体区域的包装情况。没有证据表明对这些区域有明显的选择性。如果野生型菌株168含有自主复制质粒,特别是pC194,丝裂霉素C会诱导常驻质粒DNA的大小增加,然后这些DNA会以13 kb的片段形式包装到噬菌体头部。在缺乏大部分PBSX常驻噬菌体区域的RB1144菌株中,丝裂霉素C处理不会影响常驻质粒的结构。PBSX的诱导启动了质粒上的滚环复制,然后这些质粒被包装成13 kb的片段。丝裂霉素C处理导致的质粒复制的这种改变或自噬作用,其功能需要RB1144菌株原噬菌体缺失区域内的一些DNA。