Landfear S M, Lefebvre P, Chung S, Lodish H F
Mol Cell Biol. 1982 Nov;2(11):1417-26. doi: 10.1128/mcb.2.11.1417-1426.1982.
During development of the cellular slime mold Dictyostelium discoideum, approximately 2,000 to 3,000 regulated mRNAs are induced when amoebae enter multicellular aggregates. We used in vitro transcription in isolated nuclei to follow the synthesis of individual mRNA precursors during development; these were quantitated by hybridization to cloned cDNAs or genomic DNAs. Those RNAs that are present at all stages of development--the common RNAs--were transcribed by nuclei from cells at all stages of development. By contrast, those RNAs that are present only after cells begin to aggregate--here called aggregation stage RNAs--were transcribed only by nuclei from cells at the aggregation and postaggregation stages of development. The temporal pattern of in vitro transcription correlated well with the time course of accumulation of different aggregation stage mRNAs. Continued expression of aggregation stage genes normally depends upon cell-to-cell contact or cyclic AMP (cAMP); when cells are disaggregated, the regulated mRNAs are rapidly and specifically degraded. When cAMP is subsequently added to the disaggregated cells, most of the mRNAs reaccumulate. We show here that disaggregation reduced 2- to 10-fold the relative transcription of several aggregation stage RNAs, whereas addition of cAMP to disaggregated cells reinduced the level of regulated gene transcription to values approximating those found in normal postaggregation cells. These results indicate that a representative set of Dictyostelium aggregation stage genes are under transcriptional control; both the transcription and the stability of these mRNAs require either continued cell-to-cell interactions or cAMP.
在细胞黏菌盘基网柄菌的发育过程中,当变形虫进入多细胞聚集体时,大约有2000到3000种受调控的mRNA被诱导产生。我们利用分离细胞核中的体外转录来追踪发育过程中单个mRNA前体的合成;通过与克隆的cDNA或基因组DNA杂交对这些前体进行定量。那些在发育的所有阶段都存在的RNA——即常见RNA——由处于发育各个阶段的细胞核转录。相比之下,那些仅在细胞开始聚集后才出现的RNA——这里称为聚集阶段RNA——仅由处于发育聚集阶段和聚集后阶段的细胞核转录。体外转录的时间模式与不同聚集阶段mRNA积累的时间进程密切相关。聚集阶段基因的持续表达通常依赖于细胞间接触或环磷酸腺苷(cAMP);当细胞解聚时,受调控的mRNA会迅速且特异性地降解。随后当向解聚细胞中添加cAMP时,大多数mRNA会重新积累。我们在此表明,解聚使几种聚集阶段RNA的相对转录降低了2至10倍,而向解聚细胞中添加cAMP可将受调控基因转录水平重新诱导至接近正常聚集后细胞中的水平。这些结果表明,一组具有代表性的盘基网柄菌聚集阶段基因处于转录控制之下;这些mRNA的转录和稳定性都需要持续的细胞间相互作用或cAMP。