Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Biológicas, Universitat de València, Burjassot, Spain.
PLoS One. 2010 Nov 16;5(11):e15442. doi: 10.1371/journal.pone.0015442.
The amount of mRNA in a cell is the result of two opposite reactions: transcription and mRNA degradation. These reactions are governed by kinetics laws, and the most regulated step for many genes is the transcription rate. The transcription rate, which is assumed to be exercised mainly at the RNA polymerase recruitment level, can be calculated using the RNA polymerase densities determined either by run-on or immunoprecipitation using specific antibodies. The yeast Saccharomyces cerevisiae is the ideal model organism to generate a complete set of nascent transcription rates that will prove useful for many gene regulation studies. By combining genomic data from both the GRO (Genomic Run-on) and the RNA pol ChIP-on-chip methods we generated a new, more accurate nascent transcription rate dataset. By comparing this dataset with the indirect ones obtained from the mRNA stabilities and mRNA amount datasets, we are able to obtain biological information about posttranscriptional regulation processes and a genomic snapshot of the location of the active transcriptional machinery. We have obtained nascent transcription rates for 4,670 yeast genes. The median RNA polymerase II density in the genes is 0.078 molecules/kb, which corresponds to an average of 0.096 molecules/gene. Most genes have transcription rates of between 2 and 30 mRNAs/hour and less than 1% of yeast genes have >1 RNA polymerase molecule/gene. Histone and ribosomal protein genes are the highest transcribed groups of genes and other than these exceptions the transcription of genes is an infrequent phenomenon in a yeast cell.
细胞中 mRNA 的含量是转录和 mRNA 降解两个相反反应的结果。这些反应受动力学规律的支配,许多基因最受调控的步骤是转录速率。转录速率,假定主要在 RNA 聚合酶募集水平上进行,可以使用通过特定抗体的运行或免疫沉淀确定的 RNA 聚合酶密度来计算。酵母酿酒酵母是生成完整的新生转录率集的理想模型生物,这将对许多基因调控研究非常有用。通过将 GRO(基因组运行)和 RNA pol ChIP-on-chip 方法的基因组数据结合起来,我们生成了一个新的、更准确的新生转录率数据集。通过将这个数据集与从 mRNA 稳定性和 mRNA 量数据集间接获得的数据集进行比较,我们能够获得关于转录后调控过程的生物学信息和活跃转录机制的基因组快照。我们已经获得了 4670 个酵母基因的新生转录率。基因中 RNA 聚合酶 II 的中位数密度为 0.078 个分子/kb,相当于平均每个基因有 0.096 个分子。大多数基因的转录率在 2 到 30 个 mRNA/小时之间,不到 1%的酵母基因有 >1 个 RNA 聚合酶分子/基因。组蛋白和核糖体蛋白基因是转录最高的基因群,除了这些例外,基因的转录在酵母细胞中是一种罕见的现象。