Department of Biochemistry and Biophysics, Texas A&M University, College Station, Texas, United States of America.
PLoS One. 2010 Apr 20;5(4):e10248. doi: 10.1371/journal.pone.0010248.
Induction of gene expression is correlated with alterations in nuclear organization, including proximity to other active genes, to the nuclear cortex, and to cytologically distinct domains of the nucleus. Chromosomes are tethered to the insoluble nuclear scaffold/matrix through interaction with Scaffold/Matrix Attachment Region (SAR/MAR) binding proteins. Identification and characterization of proteins involved in establishing or maintaining chromosome-scaffold interactions is necessary to understand how the nucleus is organized and how dynamic changes in attachment are correlated with alterations in gene expression. We identified and characterized one such scaffold attachment factor, a Drosophila homolog of mammalian SAF-B. The large nuclei and chromosomes of Drosophila have allowed us to show that SAF-B inhabits distinct subnuclear compartments, forms weblike continua in nuclei of salivary glands, and interacts with discrete chromosomal loci in interphase nuclei. These interactions appear mediated either by DNA-protein interactions, or through RNA-protein interactions that can be altered during changes in gene expression programs. Extraction of soluble nuclear proteins and DNA leaves SAF-B intact, showing that this scaffold/matrix-attachment protein is a durable component of the nuclear matrix. Together, we have shown that SAF-B links the nuclear scaffold, chromosomes, and transcriptional activity.
基因表达的诱导与核组织的改变有关,包括与其他活性基因、核皮质和核中细胞学上不同的结构域的接近。染色体通过与支架/基质附着区(SAR/MAR)结合蛋白相互作用而被束缚在不溶性核支架/基质上。鉴定和表征参与建立或维持染色体-支架相互作用的蛋白质对于理解核的组织方式以及附着的动态变化与基因表达的改变如何相关是必要的。我们鉴定和表征了一种这样的支架附着因子,即哺乳动物 SAF-B 的果蝇同源物。果蝇的大核和染色体使我们能够证明 SAF-B 占据不同的亚核区室,在唾液腺细胞核中形成网状连续体,并在有丝分裂核中与离散的染色体位点相互作用。这些相互作用似乎通过 DNA-蛋白质相互作用或通过 RNA-蛋白质相互作用来介导,这些相互作用在基因表达程序的改变期间可以发生改变。可溶性核蛋白和 DNA 的提取使 SAF-B 保持完整,表明这种支架/基质附着蛋白是核基质的持久组成部分。总的来说,我们已经表明 SAF-B 连接核支架、染色体和转录活性。