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酿酒酵母表现出一种孢子形成特异性的转录本积累时间模式。

Saccharomyces cerevisiae exhibits a sporulation-specific temporal pattern of transcript accumulation.

作者信息

Kaback D B, Feldberg L R

出版信息

Mol Cell Biol. 1985 Apr;5(4):751-61. doi: 10.1128/mcb.5.4.751-761.1985.

Abstract

Cultures of the yeast Saccharomyces cerevisiae that are heterozygous for the mating type (MATa/MAT alpha) undergo synchronous meiosis and spore formation when starved for nitrogen and supplied with a nonfermentable carbon source such as acetate. Haploid and homozygous MAT alpha/MAT alpha and MATa/MATa diploid cells incubated under the same conditions fail to undergo meiosis and are asporogenous. It has not yet been firmly established that gene expression during sporulation is controlled at the level of transcript accumulation. To examine this question, we used cloned genes that encode a variety of "housekeeping" functions to probe Northern blots to assay the appearance of specific transcripts in both sporulating and asporogenous S. cerevisiae. In sporulating cells, each transcript showed a characteristic pattern of accumulation, reaching a maximum relative abundance at one of several different periods. In contrast, in both asporogenous haploid MATa and diploid MAT alpha/MAT alpha cells, all transcripts accumulated with similar kinetics. These results suggest a sporulation-specific pattern for transcript appearance. During these studies, high levels of several different transcripts were observed at unexpected times in sporulating cells. Histone (H)2A and (H)2B1 transcripts, although most abundant during premeiotic DNA synthesis, remained at one-third to one-half maximal levels after its end and were found in mature ascospores. Their appearance at this time is in sharp contrast to vegetative cells in which these histone transcripts are only found just before and during the period of DNA synthesis. Furthermore, transcripts from GAL10 and CDC10 genes, which are believed to be dispensable for sporulation, were much more abundant in sporulating cells than in asporogenous cells and vegetative cells grown on glucose or acetate. The presence of these transcripts did not appear to be due to a general activation of transcription because each accumulated with different kinetics. In addition, the transcript for at least one gene, HO, that is also dispensable for sporulation was not detected. The increased abundance of transcripts from some genes not required for sporulation leads us to propose that genes preferentially expressed during sporulation need not be essential for this differentiation.

摘要

对于交配型(MATa/MATα)杂合的酿酒酵母培养物,当缺乏氮源并供应不可发酵的碳源(如乙酸盐)时,会进行同步减数分裂和孢子形成。在相同条件下培养的单倍体以及纯合的MATα/MATα和MATa/MATa二倍体细胞无法进行减数分裂,且不产生孢子。目前尚未确凿证实孢子形成过程中的基因表达是在转录本积累水平上受到调控的。为了研究这个问题,我们使用了编码各种“管家”功能的克隆基因来探测Northern印迹,以检测在进行孢子形成和不产生孢子的酿酒酵母中特定转录本的出现情况。在进行孢子形成的细胞中,每个转录本都呈现出一种特征性的积累模式,在几个不同时期中的某一个时期达到最大相对丰度。相比之下,在不产生孢子的单倍体MATa和二倍体MATα/MATα细胞中,所有转录本的积累动力学相似。这些结果表明转录本出现存在孢子形成特异性模式。在这些研究过程中,在进行孢子形成的细胞中,在一些意想不到的时间观察到了几种不同转录本的高水平表达。组蛋白(H)2A和(H)2B1转录本虽然在减数分裂前的DNA合成过程中最为丰富,但在其结束后仍保持在最大水平的三分之一到二分之一,并且在成熟的子囊中孢子中也能发现。它们此时的出现与营养细胞形成鲜明对比,在营养细胞中这些组蛋白转录本仅在DNA合成之前及合成期间被发现。此外,据信对于孢子形成可有可无的GAL10和CDC10基因的转录本,在进行孢子形成的细胞中比在不产生孢子的细胞以及在葡萄糖或乙酸盐上生长的营养细胞中丰富得多。这些转录本的存在似乎并非由于转录的普遍激活,因为它们各自以不同的动力学积累。此外,至少一个对于孢子形成也可有可无的基因HO的转录本未被检测到。一些对于孢子形成并非必需的基因转录本丰度增加,这使我们提出在孢子形成过程中优先表达的基因不一定对于这种分化是必不可少的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/10b1/366779/b87446f3178b/molcellb00100-0168-a.jpg

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