Fan Hua-Ying, Trotter Kevin W, Archer Trevor K, Kingston Robert E
Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.
Mol Cell. 2005 Mar 18;17(6):805-15. doi: 10.1016/j.molcel.2005.02.024.
SWI/SNF- and ISWI-based complexes have distinct yet overlapping chromatin-remodeling activities in vitro and perform different roles in vivo. This leads to the hypothesis that the distinct remodeling functions of these complexes are specifically required for distinct biological tasks. By creating and characterizing chimeric proteins of BRG1 and SNF2h, the motor proteins of human SWI/SNF- and ISWI-based complexes, respectively, we found that a region that includes the ATPase domain specifies the outcome of the remodeling reaction in vitro. A chimeric protein based on BRG1 but containing the SNF2h ATPase domain formed an intact SWI/SNF complex that remodeled like SNF2h. This altered-function complex was active for remodeling and could stimulate expression from some, but not all, SWI/SNF responsive promoters in vivo. Thus, we were able to separate domains of BRG1 responsible for function from those responsible for SWI/SNF complex formation and demonstrate that remodeling functions are not interchangeable in vivo.
基于SWI/SNF和ISWI的复合物在体外具有不同但重叠的染色质重塑活性,并且在体内发挥不同的作用。这引发了一种假设,即这些复合物不同的重塑功能是不同生物学任务所特需的。通过分别构建和表征人类基于SWI/SNF和ISWI的复合物的动力蛋白BRG1和SNF2h的嵌合蛋白,我们发现包含ATP酶结构域的区域在体外决定了重塑反应的结果。一种基于BRG1但含有SNF2h ATP酶结构域的嵌合蛋白形成了一个完整的SWI/SNF复合物,其重塑方式类似于SNF2h。这种功能改变的复合物具有重塑活性,并且在体内能够刺激一些(但不是全部)SWI/SNF反应性启动子的表达。因此,我们能够将BRG1中负责功能的结构域与负责SWI/SNF复合物形成的结构域区分开来,并证明重塑功能在体内是不可互换的。