Singleton C K, Manning S S, Feng Y
Department of Molecular Biology, Vanderbilt University, Nashville, Tennessee 37235.
Mol Cell Biol. 1988 Jan;8(1):10-6. doi: 10.1128/mcb.8.1.10-16.1988.
Several genes which are deactivated on the initiation of development of Dictyostelium discoideum were identified by differential screening of various cDNA libraries. These genes have in common a decrease in the steady-state levels of their corresponding mRNAs on the onset of development and as development proceeds. When development was carried out in the absence of protein synthesis by inhibition with cycloheximide, the decrease in mRNA levels for most genes (V genes) was normal or slightly accelerated. For about 5% of the genes (H genes), however, cycloheximide caused an apparent induction of expression, as revealed by a slight or dramatic increase in mRNA levels, instead of the normal decrease. This effect was due to inhibition of protein synthesis and not to cycloheximide per se. The induction was found to be due to an enhancement of the transcription rate; normal rates of transcription for the H genes were dependent on continued protein synthesis during vegetative growth and development. Thus, two general regulatory classes exist for deactivation of gene expression on initiation of development, one of which is dependent on and one of which is independent of protein synthesis. Analysis of expression of these genes in mutant strains which are aggregation deficient allowed the classes to be subdivided further. Taken together, these characterizations allow several distinct regulatory mechanisms to be identified that are involved in the deactivation of gene expression on the onset of development in D. discoideum.
通过对不同cDNA文库进行差异筛选,鉴定出了几种在盘基网柄菌发育起始时失活的基因。这些基因的共同特点是,在发育开始时以及发育进行过程中,其相应mRNA的稳态水平会降低。当通过用环己酰亚胺抑制蛋白质合成来进行发育时,大多数基因(V基因)的mRNA水平下降正常或略有加速。然而,对于约5%的基因(H基因),环己酰亚胺导致了明显的表达诱导,表现为mRNA水平轻微或显著增加,而不是正常下降。这种效应是由于蛋白质合成的抑制,而不是环己酰亚胺本身。发现这种诱导是由于转录速率的提高;H基因的正常转录速率依赖于营养生长和发育过程中持续的蛋白质合成。因此,在发育起始时基因表达失活存在两种一般调控类别,其中一种依赖于蛋白质合成,另一种不依赖于蛋白质合成。对这些基因在聚集缺陷突变菌株中的表达分析使得这些类别能够进一步细分。综上所述,这些特征使得能够鉴定出几种不同的调控机制,它们参与了盘基网柄菌发育起始时基因表达的失活。