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首页> 外文期刊>Molecular and Cellular Biology >Transcription interferes with elements important for chromosome maintenance in Saccharomyces cerevisiae.
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Transcription interferes with elements important for chromosome maintenance in Saccharomyces cerevisiae.

机译：转录干扰酿酒酵母中对于染色体维持重要的元素。

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Transcription directed into a Saccharomyces cerevisiae autonomously replicating sequence (ARS) causes high-frequency loss of minichromosomes. Conditionally stable artificial yeast chromosomes were constructed that contain an inducible GAL promoter upstream of ARS1. Under growth conditions in which the promoter was inactive, these chromosomes were mitotically stable; however, when the GAL promoter was induced, the chromosomes became extremely unstable as a result of transcriptional impairment of ARS function. This interference by the GAL promoter occurred only in cis but can occur from either side of ARS1. Transcriptional interference of ARS function can be monitored readily by using a visual colony-color assay (P. Hieter, C. Mann, M. Snyder, and R.W. Davis, Cell 40:381-392, 1985), which was further developed as a sensitive in vivo assay for sequences which rescue ARS from transcription. DNA fragments from the 3' ends of genes, inserted downstream of the GAL promoter, protected ARS function from transcriptional interference. This assay is expected to be independent of both RNA transcript stability and processing. Philippsen et al. have shown that transcription into a yeast centromere inhibits CEN function in vivo (L. Panzeri, I. Groth-Clausen, J. Shepard, A. Stotz, and P. Philippsen, Chromosomes Today 8:46-58, 1984). We identified two 200- to 300-base-pair DNA fragments flanking CEN4 that rescued ARS1 from transcription. Both of these fragments protected ARS from transcription when inserted in either orientation. The 3' ends of stable transcripts are encoded by fragments that protected the ARS from transcription, suggesting that the protection was achieved by transcription termination. It is suggested that protection of elements important for the replication and segregation of eucaryotic chromosomes from transcription is necessary for their proper function in vivo.
机译：定向到啤酒酵母自主复制序列（ARS）中的转录会导致微型染色体的高频丢失。构建条件稳定的人工酵母染色体，该染色体包含ARS1上游的可诱导GAL启动子。在启动子失活的生长条件下，这些染色体是有丝分裂稳定的。然而，当诱导GAL启动子时，由于ARS功能的转录损伤，染色体变得极为不稳定。 GAL启动子的这种干扰仅在顺式中发生，但可以在ARS1的任一侧发生。 ARS功能的转录干扰可以通过使用视觉菌落颜色测定法（P. Hieter，C. Mann，M. Snyder和RW Davis，Cell 40：381-392，1985）进行容易地监测，该方法被进一步开发为灵敏的体内测定，用于从转录中拯救ARS的序列。插入到GAL启动子下游的基因3'末端的DNA片段保护ARS功能免受转录干扰。预期该测定独立于RNA转录物稳定性和加工。 Philippsen等。已经证明转录成酵母着丝粒在体内会抑制CEN功能（L. Panzeri，I。Groth-Clausen，J。Shepard，A。Stotz，和P. Philippsen，Chromosomes Today 8：46-58，1984）。我们鉴定了两个200到300个碱基对的DNA片段，位于CEN4的两侧，这些片段从转录中拯救了ARS1。当以任一方向插入时，这两个片段都保护ARS免于转录。稳定转录本的3'端由保护ARS免受转录影响的片段编码，表明该保护作用是通过转录终止来实现的。建议保护对真核染色体的复制和分离重要的元件免受转录是其体内适当功能所必需的。

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