Brock Amy, Huang Sui, Ingber Donald E
Vascular Biology Program, Department of Pathology, Harvard Medical School and Children's Hospital, Enders 1007, 300 Longwood Ave, Boston, MA 02115, USA.
BMC Cell Biol. 2003 Jul 8;4:6. doi: 10.1186/1471-2121-4-6.
Interactions between mRNA and the cytoskeleton are critical for the localization of a number of transcripts in eukaryotic somatic cells. To characterize additional transcripts that may be subject to this form of regulation, we developed a two-step approach that utilizes biochemical fractionation of cells to isolate transcripts from different subcellular compartments followed by microarray analysis to examine and compare these subpopulations of transcripts in a massively-parallel manner.
Using this approach, mRNA was extracted from the cytoskeleton-rich and the cytosolic fractions of the promyelocytic HL-60 cell line. We identify a subset of 22 transcripts that are significantly enriched in the cytoskeleton-associated population. The majority of these encode structural proteins and/or proteins known to interact with elements of the cytoskeleton. Localization required an intact actin cytoskeleton and was largely conserved upon differentiation of precursor HL-60 cells to a macrophage-like phenotype.
We conclude that the association of transcripts with the actin cytoskeleton in somatic cells may be a critical post-transcriptional regulatory event that controls a larger class of genes than has previously been recognized.
在真核体细胞中，信使核糖核酸（mRNA）与细胞骨架之间的相互作用对于许多转录本的定位至关重要。为了鉴定可能受这种调控形式影响的其他转录本，我们开发了一种两步法，该方法利用细胞的生化分级分离从不同亚细胞区室中分离转录本，随后进行微阵列分析，以大规模平行的方式检查和比较这些转录本子群体。
使用这种方法，从早幼粒细胞HL-60细胞系富含细胞骨架和胞质的组分中提取了mRNA。我们鉴定出一组22个转录本，它们在与细胞骨架相关的群体中显著富集。其中大多数编码结构蛋白和/或已知与细胞骨架成分相互作用的蛋白质。定位需要完整的肌动蛋白细胞骨架，并且在前体HL-60细胞分化为巨噬细胞样表型时在很大程度上得以保留。
我们得出结论，体细胞中转录本与肌动蛋白细胞骨架的关联可能是一种关键的转录后调控事件，它控制的基因类别比以前认识到的更大。
