Feng G S, Chong K, Kumar A, Williams B R
Research Institute, Hospital for Sick Children, Toronto, Canada.
Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5447-51. doi: 10.1073/pnas.89.12.5447.
The double-stranded RNA (dsRNA)-binding domain of the human p68 kinase has been localized to the N-terminal half of the enzyme by using progressive deletion analysis and in vitro binding assays. To further define the domains responsible for binding to dsRNA, we cloned the mouse dsRNA-activated p65 kinase and used sequence alignment to identify conserved domains in the N-terminal region. Deletions in either of two 12-amino-acid-long and arginine- or lysine-rich regions abrogated binding to dsRNA. Moreover, in an in vivo growth inhibition assay in the yeast Saccharomyces cerevisiae, these mutants failed to exhibit a slow-growth phenotype.
通过进行渐进缺失分析和体外结合试验，已将人p68激酶的双链RNA（dsRNA）结合结构域定位到该酶的N端一半区域。为了进一步确定负责与dsRNA结合的结构域，我们克隆了小鼠dsRNA激活的p65激酶，并利用序列比对来鉴定N端区域中的保守结构域。两个富含精氨酸或赖氨酸的12个氨基酸长的区域中任一个发生缺失，都会消除与dsRNA的结合。此外，在酿酒酵母的体内生长抑制试验中，这些突变体未能表现出生长缓慢的表型。
