Shpakovskiĭ G V, Berlin Iu A
Bioorg Khim. 1983 May;9(5):711-3.
A recombination site in the transducing bacteriophage lambda plac5 DNA has been structurally elucidated. Comparison of primary structures of E. coli lac-operon (distal end of lacZ gene, Z-Y spacer, and proximal end of lacY gene) described earlier with corresponding segments of bacteriophages lambda CI857 and lambda plac 5-2 DNAs sequenced in this paper showed that the bacterial DNA insert ends immediately after Z-Y spacer, just before the initiating triplet ATG of lacY gene. It thus follows that in contrast to the earlier conception, the insert does not seem to include any part of lacY gene. The recombination sites in both phage and bacterial DNA contain structurally homological segments about 20 b. p. long (crossover region), with two extra basepairs in the bacterial DNA (AT in the sense-strand). We suppose that the very dinucleotide plays a substantial role in initiation of recombinational event: causing formation of a nonperfect heteroduplex structure, it determines the T-A internucleotide bond to be endonucleolytically cut (crossover point) followed by exonucleolytic elimination of the extra links (AT) and reciprocal strand exchange. The second recombination site in lambda plac5 DNA has been localized by us within lacI gene as being close to the HindII site (nucleotides 854 to 859 of the gene). The structures of the two regions of site-specific recombination may shed light upon mechanisms of the phage abnormal excision leading to formation of transducing phages.
转导噬菌体λplac5 DNA中的一个重组位点已在结构上得到阐明。将先前描述的大肠杆菌乳糖操纵子(lacZ基因远端、Z-Y间隔区和lacY基因近端)的一级结构与本文测序的噬菌体λCI857和λplac 5-2 DNA的相应片段进行比较,结果表明,细菌DNA插入片段在Z-Y间隔区之后、lacY基因起始三联体ATG之前立即终止。因此可以得出结论,与先前的概念相反,插入片段似乎不包括lacY基因的任何部分。噬菌体和细菌DNA中的重组位点都包含约20个碱基对长的结构同源片段(交叉区域),细菌DNA中有两个额外的碱基对(有义链中的AT)。我们推测,正是这一二核苷酸在重组事件的起始中起着重要作用:它导致形成一个非完美的异源双链结构,决定T-A核苷酸键被内切核酸酶切割(交叉点),随后通过外切核酸酶消除额外的连接(AT)并进行相互的链交换。我们已将λplac5 DNA中的第二个重组位点定位在lacI基因内,靠近HindII位点(该基因的核苷酸854至859)。位点特异性重组的这两个区域的结构可能有助于揭示导致转导噬菌体形成的噬菌体异常切除机制。