Farrar N A, Kiyosawa H, Hughes J E, Welker D L, Williams K L
School of Biological Sciences, Macquarie University, Sydney, New South Wales, Australia.
Plasmid. 1994 Mar;31(2):184-95. doi: 10.1006/plas.1994.1019.
The complete nucleotide sequence of Ddp1, a high copy number 13.7-kbp endogenous nuclear plasmid of Dictyostelium discoideum is presented. Previous studies have shown nine transcripts which map to five different regions of Ddp1, suggesting alternative transcription initiation sites and/or post-transcriptional processing. The sequence presented here shows five long open reading frames corresponding to previously known transcribed regions, as well as an additional reading frame, in the region of the presumed origin of replication. Two of the predicted proteins from the Ddp1 reading frames have potential leader sequences and transmembrane domains, while the rest, if translated, would encode soluble proteins. One of these has both leucine zipper and zinc finger-like motifs. Sequences upstream of the reading frames show strong similarities to chromosomal promoter elements, suggesting that transcription of at least some of the plasmid-encoded genes is regulated by factors which regulate chromosomal transcription in this organism. The region containing the presumptive origin of replication contains the promoters of the major growth-specific gene, as well as a second gene; thus, the processes of transcription and replication of Ddp1 may be intimately linked. The origin-encompassing region has a long homopurine/homopyrimidine stretch, which is similar to sequences shown in vitro to be capable of forming a triple helix structure through Hoogsteen base pairing. Possible roles for these sequences and the products of the ORFs in plasmid maintenance are discussed.
本文给出了盘基网柄菌(Dictyostelium discoideum)中一个高拷贝数的13.7kbp内源性核质粒Ddp1的完整核苷酸序列。先前的研究已显示有9种转录本,它们定位于Ddp1的5个不同区域,这表明存在不同的转录起始位点和/或转录后加工过程。这里给出的序列显示有5个长开放阅读框,对应于先前已知的转录区域,以及在假定的复制起点区域的一个额外阅读框。Ddp1阅读框预测的两种蛋白质具有潜在的前导序列和跨膜结构域,而其余的如果被翻译,则将编码可溶性蛋白质。其中一种蛋白质同时具有亮氨酸拉链和锌指样基序。阅读框上游的序列与染色体启动子元件有很强的相似性,这表明至少一些质粒编码基因的转录受调控该生物体染色体转录的因子调控。包含假定复制起点的区域含有主要生长特异性基因以及另一个基因的启动子;因此,Ddp1的转录和复制过程可能紧密相连。包含起点的区域有一段长的同型嘌呤/同型嘧啶序列,类似于体外显示能够通过Hoogsteen碱基配对形成三螺旋结构的序列。文中讨论了这些序列和开放阅读框产物在质粒维持中的可能作用。