A novel plasmid-based microarray screen identifies suppressors of rrp6Delta in Saccharomyces cerevisiae
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 rrp6Delta...
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| Veröffentlicht in: | Molecular and cellular biology 2007-02, Vol.27 (3), p.1044 |
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| creator | Abruzzi, Katharine Denome, Sylvia Olsen, Jens Raabjerg Assenholt, Jannie Haaning, Line Lindegaard Jensen, Torben Heick Rosbash, Michael |
| description | 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 rrp6Delta 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 rrp6Delta strains at 37 degrees 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 rrp6Delta strains. Microarray analyses of gene expression in rrp6Delta strains and a number of suppressor strains support this hypothesis. |
| format | Article |
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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 rrp6Delta 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 rrp6Delta strains at 37 degrees 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 rrp6Delta strains. 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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 rrp6Delta 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 rrp6Delta strains at 37 degrees 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 rrp6Delta strains. Microarray analyses of gene expression in rrp6Delta strains and a number of suppressor strains support this hypothesis.</description><subject>Down-Regulation</subject><subject>Exoribonucleases - genetics</subject><subject>Exoribonucleases - metabolism</subject><subject>Exosome Multienzyme Ribonuclease Complex</subject><subject>Gene Deletion</subject><subject>Gene Expression Regulation, Fungal - genetics</subject><subject>Genes, Fungal - genetics</subject><subject>Genes, Suppressor</subject><subject>Oligonucleotide Array Sequence Analysis - methods</subject><subject>Plasmids - genetics</subject><subject>Polyadenylation</subject><subject>Protein Binding</subject><subject>RNA Stability</subject><subject>RNA, Fungal - genetics</subject><subject>RNA, Fungal - metabolism</subject><subject>RNA, Messenger - genetics</subject><subject>RNA, Messenger - metabolism</subject><subject>RNA, Ribosomal - genetics</subject><subject>RNA, Ribosomal - metabolism</subject><subject>RNA, Small Nucleolar - genetics</subject><subject>RNA, Small Nucleolar - metabolism</subject><subject>RNA-Binding Proteins - metabolism</subject><subject>Saccharomyces cerevisiae - cytology</subject><subject>Saccharomyces cerevisiae - genetics</subject><subject>Saccharomyces cerevisiae - growth & development</subject><subject>Saccharomyces cerevisiae Proteins - genetics</subject><subject>Saccharomyces cerevisiae Proteins - metabolism</subject><subject>Suppression, Genetic</subject><subject>Temperature</subject><issn>0270-7306</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo1j81KAzEURrNQbK2-guQFBjKTTNJZlvoLBRd2X25ybzQyP-GmLczbW1BXHwcOB74rsVSNU5XTyi7EbSnfSinbKX0jFrWrVe2cWYrPjRynM_Uy91CGhJWHQiiHFHgCZphlCUw0yoQ0HlNMVGQ55cxUysRFTlEyZ_tI_RFkGuUHhPAFPA1zuJiBmM6pJKA7cR2hL3T_tyuxf37ab1-r3fvL23azq3JrTEVoqXEeNay7NUJwaNDbaNoO9IUa7ExsVIsYCWPonKfGBBOt87pVOmq9Eg-_2XzyA-EhcxqA58P_Yf0DZ79UQg</recordid><startdate>200702</startdate><enddate>200702</enddate><creator>Abruzzi, Katharine</creator><creator>Denome, Sylvia</creator><creator>Olsen, Jens Raabjerg</creator><creator>Assenholt, Jannie</creator><creator>Haaning, Line Lindegaard</creator><creator>Jensen, Torben Heick</creator><creator>Rosbash, Michael</creator><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope></search><sort><creationdate>200702</creationdate><title>A novel plasmid-based microarray screen identifies suppressors of rrp6Delta in Saccharomyces cerevisiae</title><author>Abruzzi, Katharine ; Denome, Sylvia ; Olsen, Jens Raabjerg ; Assenholt, Jannie ; Haaning, Line Lindegaard ; Jensen, Torben Heick ; Rosbash, Michael</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p544-ed6e27bd3a898dac7d4db6f459a3ac72d94f205ddfedfc97be24c4f67b3503f33</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Down-Regulation</topic><topic>Exoribonucleases - genetics</topic><topic>Exoribonucleases - metabolism</topic><topic>Exosome Multienzyme Ribonuclease Complex</topic><topic>Gene Deletion</topic><topic>Gene Expression Regulation, Fungal - genetics</topic><topic>Genes, Fungal - genetics</topic><topic>Genes, Suppressor</topic><topic>Oligonucleotide Array Sequence Analysis - methods</topic><topic>Plasmids - genetics</topic><topic>Polyadenylation</topic><topic>Protein Binding</topic><topic>RNA Stability</topic><topic>RNA, Fungal - genetics</topic><topic>RNA, Fungal - metabolism</topic><topic>RNA, Messenger - genetics</topic><topic>RNA, Messenger - metabolism</topic><topic>RNA, Ribosomal - genetics</topic><topic>RNA, Ribosomal - metabolism</topic><topic>RNA, Small Nucleolar - genetics</topic><topic>RNA, Small Nucleolar - metabolism</topic><topic>RNA-Binding Proteins - metabolism</topic><topic>Saccharomyces cerevisiae - cytology</topic><topic>Saccharomyces cerevisiae - genetics</topic><topic>Saccharomyces cerevisiae - growth & development</topic><topic>Saccharomyces cerevisiae Proteins - genetics</topic><topic>Saccharomyces cerevisiae Proteins - metabolism</topic><topic>Suppression, Genetic</topic><topic>Temperature</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Abruzzi, Katharine</creatorcontrib><creatorcontrib>Denome, Sylvia</creatorcontrib><creatorcontrib>Olsen, Jens Raabjerg</creatorcontrib><creatorcontrib>Assenholt, Jannie</creatorcontrib><creatorcontrib>Haaning, Line Lindegaard</creatorcontrib><creatorcontrib>Jensen, Torben Heick</creatorcontrib><creatorcontrib>Rosbash, Michael</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><jtitle>Molecular and cellular biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Abruzzi, Katharine</au><au>Denome, Sylvia</au><au>Olsen, Jens Raabjerg</au><au>Assenholt, Jannie</au><au>Haaning, Line Lindegaard</au><au>Jensen, Torben Heick</au><au>Rosbash, Michael</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A novel plasmid-based microarray screen identifies suppressors of rrp6Delta in Saccharomyces cerevisiae</atitle><jtitle>Molecular and cellular biology</jtitle><addtitle>Mol Cell Biol</addtitle><date>2007-02</date><risdate>2007</risdate><volume>27</volume><issue>3</issue><spage>1044</spage><pages>1044-</pages><issn>0270-7306</issn><abstract>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 rrp6Delta 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 rrp6Delta strains at 37 degrees 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 rrp6Delta strains. Microarray analyses of gene expression in rrp6Delta strains and a number of suppressor strains support this hypothesis.</abstract><cop>United States</cop><pmid>17101774</pmid></addata></record> |
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| ispartof | Molecular and cellular biology, 2007-02, Vol.27 (3), p.1044 |
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| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Alma/SFX Local Collection |
| subjects | Down-Regulation Exoribonucleases - genetics Exoribonucleases - metabolism Exosome Multienzyme Ribonuclease Complex Gene Deletion Gene Expression Regulation, Fungal - genetics Genes, Fungal - genetics Genes, Suppressor Oligonucleotide Array Sequence Analysis - methods Plasmids - genetics Polyadenylation Protein Binding RNA Stability RNA, Fungal - genetics RNA, Fungal - metabolism RNA, Messenger - genetics RNA, Messenger - metabolism RNA, Ribosomal - genetics RNA, Ribosomal - metabolism RNA, Small Nucleolar - genetics RNA, Small Nucleolar - metabolism RNA-Binding Proteins - metabolism Saccharomyces cerevisiae - cytology Saccharomyces cerevisiae - genetics Saccharomyces cerevisiae - growth & development Saccharomyces cerevisiae Proteins - genetics Saccharomyces cerevisiae Proteins - metabolism Suppression, Genetic Temperature |
| title | A novel plasmid-based microarray screen identifies suppressors of rrp6Delta in Saccharomyces cerevisiae |
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