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A dynamic scaffold of Pre-snoRNP factors facilitates human box C/D snoRNP assembly

McKeegan, K. S.; Debieux, C. M.; Boulon, S.; Bertrand, E.; Watkins, N. J.

Molecular and Cellular Biology

2007-10 / vol 27 / pages 6782-6793

Abstract

The box C/D small nucleolar RNPs (snoRNPs) are essential for the processing and modification of rRNA. The core box C/D proteins are restructured during human U3 box C/D snoRNP biogenesis; however, the molecular basis of this is unclear. Here we show that the U8 snoRNP is also restructured, suggesting that this may occur with all box C/D snoRNPs. We have characterized four novel human biogenesis factors (BCD1, NOP17, NUFIP, and TAF9) which, along with the ATPases TIP48 and TIP49, are likely to be involved in the formation of the pre-snoRNP. We have analyzed the in vitro protein-protein interactions between the assembly factors and core box C/D proteins. Surprisingly, this revealed few interactions between the individual core box C/D proteins. However, the novel biogenesis factors and TIP48 and TIP49 interacted with one or more of the core box C/D proteins, implying that they mediate the assembly of the pre-snoRNP. Consistent with this, we show that NUFIP bridges interactions between the core box C/D proteins in a partially reconstituted pre-snoRNP. Restructuring of the core complex probably reflects the conversion of the pre-snoRNP, where core protein-protein interactions are maintained by the bridging biogenesis factors, to the mature snoRNP.

0270-7306

Tags

mammalian-cells; transcription; small nucleolar rnas; polymerase-ii; chromatin remodeling complex; noncoding rnas; protein-protein; ribosome biogenesis; tri-snrnp; u3 snornp

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