Sarnowski Tomasz J, Swiezewski Szymon, Pawlikowska Katarzyna, Kaczanowski Szymon, Jerzmanowski Andrzej
Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw University, Pawińskiego 5A, 02-106 Warsaw, Poland.
Nucleic Acids Res. 2002 Aug 1;30(15):3412-21. doi: 10.1093/nar/gkf458.
ATP-dependent nucleosome remodeling plays a central role in the regulation of access to chromatin DNA. Swi/Snf remodeling complexes characterized in yeast, Drosophila and mammals all contain a conserved set of core subunits composed of homologs of yeast SNF2-type DNA-dependent ATPase, SNF5 and SWI3 proteins. So far, no complete Swi/Snf-type complex has been characterized in plants. Arabidopsis contains a single SNF5-type gene, BSH, which has been shown to complement the yeast snf5 mutation. Here we describe the characterization of AtSWI3B, the smallest of the four Arabidopsis homologs of SWI3. The gene encoding AtSWI3B is expressed ubiquitously in the plant. AtSWI3B is localized to nuclei and is associated mostly with the chromatin and soluble protein fractions. When expressed in Saccharomyces cerevisiae, the cDNA encoding AtSWI3B partially complements the swi3 mutant phenotype. However, like BSH, AtSWI3B is unable to activate transcription in yeast when tethered to DNA. The analysis by yeast two-hybrid indicates that AtSWI3B is capable of forming homodimers and interacts with BSH as well as with two other members of the Arabidopsis SWI3 family: AtSWI3A and AtSWI3C. The results of phage display screen using recombinant protein, confirmed by direct yeast two-hybrid analyses, indicate that AtSWI3B interacts with FCA, a regulator of flowering time in Arabidopsis. This interaction is through the C-terminal region of FCA, located outside the conserved RNA- and protein-binding domains of this protein.
ATP 依赖的核小体重塑在调控染色质 DNA 的可及性方面发挥着核心作用。在酵母、果蝇和哺乳动物中鉴定出的 Swi/Snf 重塑复合物均包含一组保守的核心亚基,这些亚基由酵母 SNF2 型 DNA 依赖的 ATP 酶、SNF5 和 SWI3 蛋白的同源物组成。到目前为止,尚未在植物中鉴定出完整的 Swi/Snf 型复合物。拟南芥含有一个单一的 SNF5 型基因 BSH,该基因已被证明可弥补酵母 snf5 突变。在此,我们描述了 AtSWI3B 的特性,它是拟南芥中 SWI3 的四个同源物中最小的一个。编码 AtSWI3B 的基因在植物中普遍表达。AtSWI3B 定位于细胞核,并且主要与染色质和可溶性蛋白组分相关。当在酿酒酵母中表达时,编码 AtSWI3B 的 cDNA 部分弥补了 swi3 突变体表型。然而,与 BSH 一样,当与 DNA 相连时,AtSWI3B 在酵母中无法激活转录。酵母双杂交分析表明,AtSWI3B 能够形成同二聚体,并与 BSH 以及拟南芥 SWI3 家族的另外两个成员 AtSWI3A 和 AtSWI3C 相互作用。使用重组蛋白进行的噬菌体展示筛选结果,经直接酵母双杂交分析证实,表明 AtSWI3B 与 FCA 相互作用,FCA 是拟南芥开花时间的调节因子。这种相互作用是通过 FCA 的 C 末端区域,该区域位于该蛋白保守的 RNA 和蛋白结合结构域之外。