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1A6/DRIM, the human UTP20 functions in 28S and 5.8S rRNA processing

1A6/DRIM, the human UTP20 functions in 28S and 5.8S rRNA processing
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摘要 1A6/DRIM has been identified as UTP20, a small subunit processome component, functioning in 18S rRNA processing. In the present study, the maturation of 28S rRNA and 5.8S rRNA was inhibited when 1A6/DRIM was silenced in HeLa cells; and coincidently, an accumulation of 32S rRNA precursor was observed. Immunoprecipitation was performed with the anti-1A6/DRIM antibody, followed by Northern blot with the ITS2 probe. The results showed that 1A6/DRIM was associated with both 32S and 12S rRNA precursors in vivo. The expression profile of 1A6/DRIM during rRNA processing was investigated by sucrose density gradient fractionation in combination with Western blot analysis. The results demonstrated that 1A6/DRIM was involved in the pre-60S particles in addition to the pre-40S particles and co-sediment with the 32S and 12S rRNA precursors in the nucleolus. Furthermore, the interaction of U8 snoRNA with 1A6/DRIM was revealed by immunoprecipitation. These results demonstrated that 1A6/DRIM interacted with both 32S rRNA and U8 snoRNA, being involved in 28S rRNA and 5.8S rRNA processing. 1A6/DRIM has been identified as UTP20, a small subunit processome component, functioning in 18S rRNA processing. In the present study, the maturation of 28S rRNA and 5.8S rRNA was inhibited when 1A6/DRIM was silenced in HeLa cells; and coincidently, an accumulation of 32S rRNA precursor was observed. Immunoprecipitation was performed with the anti-lA6/DRIM antibody, followed by Northern blot with the ITS2 probe. The results showed that 1A6/DRIM was associated with both 32S and 12S rRNA precursors in vivo. The expression profile of 1A6/DRIM during rRNA processing was investigated by sucrose density gradient fractionation in combination with Western blot analysis. The results demonstrated that 1A6/DRIM was involved in the pre-60S particles in addition to the pre-40S particles and co-sediment with the 32S and 12S rRNA precursors in the nucleolus. Furthermore, the interaction of U8 snoRNA with 1A6/DRIM was revealed by immunoprecipitation. These results demonstrated that 1A6/DRIM interacted with both 32S rRNA and U8 snoRNA, being involved in 28S rRNA and 5.8S rRNA processing.
出处 《Chinese Science Bulletin》 SCIE EI CAS 2010年第17期1770-1776,共7页
基金 supported by the National High-Tech Research and Development Program of China (2006AA02A402 and 2008AA02Z131) the National Natural Science Foundation of China (30771224) the Ministry of Education 985 project of China (985-2-016-24) National Key Basic Research Program of China (2006CB943603)
关键词 RRNA基因 加工过程 SNORNA 职能 人类 HELA细胞 免疫印迹分析 免疫沉淀 1A6/DRIM, UTP20, small-subunit (SSU) processome, 28S and 5.8S rRNA processing, U8 small nucleolar RNA (U8 snoRNA)
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