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Molecular and Cellular Biology, February 2006, p. 1183-1194, Vol. 26, No. 4
0270-7306/06/$08.00+0 doi:10.1128/MCB.26.4.1183-1194.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.
Departments of Molecular Biophysics and Biochemistry,1 Genetics,2 Therapeutic Radiology, Yale University School of Medicine, New Haven, Connecticut 065203
Received 25 July 2005/ Returned for modification 19 August 2005/ Accepted 14 November 2005
The 17 putative RNA helicases required for pre-rRNA processing are predicted to play a crucial role in ribosome biogenesis by driving structural rearrangements within preribosomes. To better understand the function of these proteins, we have generated a battery of mutations in five putative RNA helicases involved in 18S rRNA synthesis and analyzed their effects on cell growth and pre-rRNA processing. Our results define functionally important residues within conserved motifs and demonstrate that lethal mutations in predicted ATP binding-hydrolysis motifs often confer a dominant negative phenotype in vivo when overexpressed in a wild-type background. We show that dominant negative mutants delay processing of the 35S pre-rRNA and cause accumulation of pre-rRNA species that normally have low steady-state levels. Our combined results establish that not all conserved domains function identically in each protein, suggesting that the RNA helicases may have distinct biochemical properties and diverse roles in ribosome biogenesis.
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