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Molecular and Cellular Biology, February 2007, p. 1044-1055, Vol. 27, No. 3
0270-7306/07/$08.00+0     doi:10.1128/MCB.01299-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

A Novel Plasmid-Based Microarray Screen Identifies Suppressors of rrp6 in Saccharomyces cerevisiae^,

Katharine Abruzzi,^1, Sylvia Denome,^1, Jens Raabjerg Olsen,^2, Jannie Assenholt,² Line Lindegaard Haaning,² Torben Heick Jensen,² and Michael Rosbash^1*

Howard Hughes Medical Institute and Department of Biology, Brandeis University, Waltham, Massachusetts 02454,¹ Centre for mRNP Biogenesis and Metabolism, Department of Molecular Biology, Aarhus University, C. F. Møllers Alle, Building 130, 8000 Aarhus C, Denmark²

Received 15 July 2006/ Returned for modification 7 August 2006/ Accepted 30 October 2006

Genetic screens in Saccharomyces cerevisiae provide novel information about interacting genes and pathways. We screened for high-copy-number suppressors of a strain with the gene encoding the nuclear exosome component Rrp6p deleted, with either a traditional plate screen for suppressors of rrp6 temperature sensitivity or a novel microarray enhancer/suppressor screening (MES) strategy. MES combines DNA microarray technology with high-copy-number plasmid expression in liquid media. The plate screen and MES identified overlapping, but also different, suppressor genes. Only MES identified the novel mRNP protein Nab6p and the tRNA transporter Los1p, which could not have been identified in a traditional plate screen; both genes are toxic when overexpressed in rrp6 strains at 37°C. Nab6p binds poly(A)⁺ RNA, and the functions of Nab6p and Los1p suggest that mRNA metabolism and/or protein synthesis are growth rate limiting in rrp6 strains. Microarray analyses of gene expression in rrp6 strains and a number of suppressor strains support this hypothesis.

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* Corresponding author. Mailing address: Howard Hughes Medical Institute and Department of Biology, Brandeis University, Waltham, MA 02454. Phone: (781) 736-3160. Fax: (781) 736-3164. E-mail: rosbash{at}brandeis.edu.

Published ahead of print on 13 November 2006.

Supplemental material for this article may be found at http://mcb.asm.org/.

K.A. and S.D. contributed equally to this work.

Present address: Interdisciplinary Nanoscience Center iNANO, University of Aarhus, 8000 Aarhus C, Denmark.

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Molecular and Cellular Biology, February 2007, p. 1044-1055, Vol. 27, No. 3
0270-7306/07/$08.00+0     doi:10.1128/MCB.01299-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

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