Emond E, Dion E, Walker S A, Vedamuthu E R, Kondo J K, Moineau S
Department of Biochemistry and Groupe de Recherche en Ecologie Buccale, Faculté de Médecine Dentaire, Université Laval, Québec, G1K 7P4 Canada.
Appl Environ Microbiol. 1998 Dec;64(12):4748-56. doi: 10.1128/AEM.64.12.4748-4756.1998.
Lactococcus lactis W-37 is highly resistant to phage infection. The cryptic plasmids from this strain were coelectroporated, along with the shuttle vector pSA3, into the plasmid-free host L. lactis LM0230. In addition to pSA3, erythromycin- and phage-resistant isolates carried pSRQ900, an 11-kb plasmid from L. lactis W-37. This plasmid made the host bacteria highly resistant (efficiency of plaquing <10(-8)) to c2- and 936-like phages. pSRQ900 did not confer any resistance to phages of the P335 species. Adsorption, cell survival, and endonucleolytic activity assays showed that pSRQ900 encodes an abortive infection mechanism. The phage resistance mechanism is limited to a 2.2-kb EcoRV/BclI fragment. Sequence analysis of this fragment revealed a complete open reading frame (abiQ), which encodes a putative protein of 183 amino acids. A frameshift mutation within abiQ completely abolished the resistant phenotype. The predicted peptide has a high content of positively charged residues (pI = 10.5) and is, in all likelihood, a cytosolic protein. AbiQ has no homology to known or deduced proteins in the databases. DNA replication assays showed that phage c21 (c2-like) and phage p2 (936-like) can still replicate in cells harboring AbiQ. However, phage DNA accumulated in its concatenated form in the infected AbiQ+ cells, whereas the AbiQ- cells contained processed (mature) phage DNA in addition to the concatenated form. The production of the major capsid protein of phage c21 was not hindered in the cells harboring AbiQ.
乳酸乳球菌W - 37对噬菌体感染具有高度抗性。将该菌株的隐蔽质粒与穿梭载体pSA3一起电穿孔导入无质粒宿主乳酸乳球菌LM0230。除了pSA3外,抗红霉素和噬菌体的分离株携带pSRQ900,这是一种来自乳酸乳球菌W - 37的11 kb质粒。该质粒使宿主细菌对c2样和936样噬菌体具有高度抗性(噬斑形成效率<10(-8))。pSRQ900对P335种噬菌体没有赋予任何抗性。吸附、细胞存活和核酸内切酶活性测定表明pSRQ900编码一种流产感染机制。噬菌体抗性机制限于一个2.2 kb的EcoRV/BclI片段。该片段的序列分析揭示了一个完整的开放阅读框(abiQ),其编码一个183个氨基酸的推定蛋白质。abiQ内的移码突变完全消除了抗性表型。预测的肽具有高含量的带正电荷残基(pI = 10.5),很可能是一种胞质蛋白。AbiQ与数据库中已知或推导的蛋白质没有同源性。DNA复制测定表明噬菌体c21(c2样)和噬菌体p2(936样)仍可在携带AbiQ的细胞中复制。然而,噬菌体DNA以其串联形式积累在感染的AbiQ +细胞中,而AbiQ -细胞除了串联形式外还含有加工后的(成熟的)噬菌体DNA。在携带AbiQ的细胞中,噬菌体c21主要衣壳蛋白的产生没有受到阻碍。