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Molecular and Cellular Biology, October 2002, p. 7258-7267, Vol. 22, No. 20
0270-7306/02/$04.00+0     DOI: 10.1128/MCB.22.20.7258-7267.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Components of an Interdependent Unit within the SSU Processome Regulate and Mediate Its Activity

Karen A. Wehner,1 Jennifer E. G. Gallagher,1 and Susan J. Baserga1,2*

Departments of Genetics,1 Therapeutic Radiology, Yale University School of Medicine, New Haven, Connecticut 06520-80402

Received 26 April 2002/ Returned for modification 13 June 2002/ Accepted 9 July 2002

The SSU processome is required for production of the small ribosomal subunit RNA, the 18S rRNA. Specifically, the U3 small nucleolar RNA (snoRNA) component of the SSU processome is essential for the formation of the conserved central pseudoknot and for cleavages of the pre-rRNA, both of which are required for 18S maturation. To further elucidate how these events are mediated, we examined the regulatory and mechanistic roles of the U3 specific proteins: Imp3p, Imp4p, and Mpp10p. We found that these proteins demonstrated an interdependence with respect to their stability and to their association with the U3 snoRNA. Because mutations in the U3 snoRNA that disrupt pre-rRNA processing confer similar defects on growth and pre-rRNA processing as do carboxy-terminal truncations of Mpp10p, we hypothesized that Mpp10p may be involved in maintaining U3 snoRNA-pre-rRNA base pairing. However, combining the two mutations resulted in a more pronounced cleavage defect at site A2, suggesting that Mpp10p is also required at an additional mechanistic step. Furthermore, heterologous complementation experiments demonstrate that the last 95 amino acids of yeast Mpp10p are specifically required for growth and pre-rRNA processing at low temperatures.

* Corresponding author. Mailing address: Departments of Genetics and Therapeutic Radiology, Yale University School of Medicine, P.O. Box 208040, New Haven, CT 06520-8040. Phone: (203) 785-4618. Fax: (203) 785-6309. E-mail: susan.baserga{at}yale.edu.

Molecular and Cellular Biology, October 2002, p. 7258-7267, Vol. 22, No. 20
0022-538X/02/$04.00+0     DOI: 10.1128/MCB.22.20.7258-7267.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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