Henning Dale, So Rolando B, Jin Runyan, Lau Lester F, Valdez Benigno C
Department of Pharmacology, Baylor College of Medicine, Houston, Texas 77030, USA.
J Biol Chem. 2003 Dec 26;278(52):52307-14. doi: 10.1074/jbc.M310846200. Epub 2003 Oct 13.
The intricate production of ribosomal RNA is well defined in yeast, but its complexity in higher organisms is barely understood. We recently showed that down-regulation of nucleolar protein RNA helicase II/Gualpha (RH-II/Gualpha or DDX21) in Xenopus oocytes inhibited processing of 20 S rRNA to 18 S and contributed to degradation of 28 S rRNA (Yang, H., Zhou, J., Ochs, R. L., Henning, D., Jin, R., and Valdez, B. C. (2003) J. Biol. Chem. 278, 38847-38859). Since no nucleolar RNA helicase has been functionally characterized in mammalian cells, we used short interfering RNA to search for functions for RH-II/Gualpha and its paralogue RH-II/Gubeta in rRNA production. Silencing of RH-II/Gualpha by more than 80% in HeLa cells resulted in an almost 80% inhibition of 18 and 28 S rRNA production. This inhibition could be reversed by exogenous expression of wild type RH-II/Gualpha. A helicase-deficient mutant form having ATPase activity was able to rescue the production of 28 S but not 18 S rRNA. A phenotype exhibiting inhibition of 18 S and 28 S rRNA production was also observed when the paralogue RH-II/Gubeta was overexpressed. Both down-regulation of RH-II/Gualpha and overexpression of RH-II/Gubeta slowed cell proliferation. The opposite effects of the two paralogues suggest antagonistic functions.
核糖体RNA复杂的产生过程在酵母中已得到充分阐明,但在高等生物中的复杂性却几乎不为人知。我们最近发现,非洲爪蟾卵母细胞中核仁蛋白RNA解旋酶II/α亚基(RH-II/α或DDX21)的下调会抑制20 S rRNA加工成18 S rRNA,并导致28 S rRNA的降解(Yang, H., Zhou, J., Ochs, R. L., Henning, D., Jin, R., and Valdez, B. C. (2003) J. Biol. Chem. 278, 38847 - 38859)。由于在哺乳动物细胞中尚未对任何核仁RNA解旋酶进行功能鉴定,我们使用小干扰RNA来探寻RH-II/α及其旁系同源物RH-II/β在rRNA产生过程中的功能。在HeLa细胞中使RH-II/α沉默超过80%,导致18 S和28 S rRNA的产生受到近80%的抑制。这种抑制可通过野生型RH-II/α的外源表达得以逆转。具有ATP酶活性的解旋酶缺陷突变体形式能够挽救28 S rRNA的产生,但不能挽救18 S rRNA的产生。当旁系同源物RH-II/β过表达时,也观察到了抑制18 S和28 S rRNA产生的表型。RH-II/α的下调和RH-II/β的过表达均减缓了细胞增殖。这两个旁系同源物的相反作用表明它们具有拮抗功能。