Wegrzyn G, Wegrzyn A, Konieczny I, Bielawski K, Konopa G, Obuchowski M, Helinski D R, Taylor K
Department of Molecular Biology, University of Gdańsk, Poland.
Genetics. 1995 Apr;139(4):1469-81. doi: 10.1093/genetics/139.4.1469.
We demonstrate that the initiation of coliphage lambda DNA replication is dependent on the host initiator function dnaA, provided that the lambdoid prophage Rac is absent. Presence of Rac compensated the absence of dnaA function, causing initiation of replication. In dnaAts rac+ cells at 43 degrees, most of parental phage DNA molecules, after one round of theta replication, switched to a replication with features of the sigma mode and produced progeny at high yield. Initiation of replication of the lambda Pts1 mutant at 43 degrees was blocked by dnaA function; however, under dnaA-rac+ conditions all parental phage DNA molecules, after one round of theta replication, switched to the sigma mode and produced progeny at high yield. Taking into account our recent finding that transcriptional activation of ori lambda seems to be dnaA-regulated (to be published elsewhere), we suggest that the DnaA-lambda Pts1 incompatibility occurs at the insertion of the ori lambda-bound lambda O-lambda P-DnaB preprimosome between the complementary lambda DNA strands. The role of Rac and the mechanism of the switch from theta to sigma mode of lambda phage DNA replication are discussed.
我们证明,只要不存在类λ原噬菌体Rac,大肠杆菌噬菌体λ DNA复制的起始就依赖于宿主起始因子功能dnaA。Rac的存在弥补了dnaA功能的缺失,从而引发复制。在43℃的dnaAts rac+细胞中,大多数亲代噬菌体DNA分子在一轮θ型复制后,转变为具有σ型模式特征的复制,并高产产生子代。λ Pts1突变体在43℃时的复制起始被dnaA功能阻断;然而,在dnaA-rac+条件下,所有亲代噬菌体DNA分子在一轮θ型复制后,都转变为σ型模式并高产产生子代。考虑到我们最近的发现,即ori λ的转录激活似乎受dnaA调控(将在其他地方发表),我们认为DnaA与λ Pts1的不相容性发生在ori λ结合的λ O-λ P-DnaB预引发体插入互补λ DNA链之间时。本文讨论了Rac的作用以及λ噬菌体DNA复制从θ型模式转变为σ型模式的机制。