Yang Hushan, Zhou Juhua, Ochs Robert L, Henning Dale, Jin Runyan, Valdez Benigno C
Department of Pharmacology, Baylor College of Medicine, Houston, Texas 77030, USA.
J Biol Chem. 2003 Oct 3;278(40):38847-59. doi: 10.1074/jbc.M302258200. Epub 2003 Jul 8.
Genetic manipulations have revealed the functions of RNA helicases in ribosomal RNA (rRNA) biogenesis in yeast. However, no report shows the role of an RNA helicase in rRNA formation in higher eukaryotes. This study reports the functional characterization of the frog homologue of nucleolar RNA helicase II/Gu (xGu or DDX21). Down-regulation of xGu in Xenopus laevis oocyte using an antisense oligodeoxynucleotide results in the depletion of 18 and 28 S rRNAs. The disappearance of 18 S rRNA is accompanied by an accumulation of 20 S, indicating that xGu is critical in the processing of 20 to 18 S rRNA. The degradation of 28 S rRNA into fragments smaller than 18 S is also associated with a specific decrease in the level of xGu protein. These effects are reversed in the presence of in vitro synthesized wild type xGu mRNA but not its helicase-deficient mutant form. Similar aberrant rRNA processing is observed when antibody against xGu is microinjected. The involvement of xGu in processing of rRNA is consistent with the localization of Gu protein to the granular and dense fibrillar components of PtK2 cell nucleoli by immunoelectron microscopy. Our results show that xGu is involved in the processing of 20 to 18 S rRNA and contributes to the stability of 28 S rRNA in Xenopus oocytes.
基因操作已揭示了RNA解旋酶在酵母核糖体RNA(rRNA)生物合成中的功能。然而,尚无报告显示RNA解旋酶在高等真核生物rRNA形成中的作用。本研究报告了核仁RNA解旋酶II/Gu(xGu或DDX21)的青蛙同源物的功能特性。使用反义寡脱氧核苷酸下调非洲爪蟾卵母细胞中的xGu会导致18和28 S rRNA的消耗。18 S rRNA的消失伴随着20 S的积累,表明xGu在20至18 S rRNA的加工中起关键作用。28 S rRNA降解为小于18 S的片段也与xGu蛋白水平的特异性降低有关。在体外合成的野生型xGu mRNA存在下,这些效应会逆转,但其解旋酶缺陷型突变体形式则不会。当显微注射抗xGu抗体时,会观察到类似的异常rRNA加工。通过免疫电子显微镜观察,xGu参与rRNA加工与Gu蛋白定位于PtK2细胞核仁的颗粒和致密纤维成分一致。我们的结果表明,xGu参与了非洲爪蟾卵母细胞中20至18 S rRNA的加工,并有助于28 S rRNA的稳定性。