Biochemical and genetic analysis of the yeast proteome with a movable ORF collection

Daniel M. Gelperin; Michael A. White; Martha L. Wilkinson; Yoshiko Kon; Li A. Kung; Kevin J. Wise; Nelson Lopez-Hoyo; Lixia Jiang; Stacy Piccirillo; Haiyuan Yu; Mark Gerstein; Mark E. Dumont; Eric M. Phizicky; Michael Snyder; Elizabeth J. Grayhack

doi:10.1101/gad.1362105

Biochemical and genetic analysis of the yeast proteome with a movable ORF collection

¹Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642, USA; ²Department of Molecular, Cellular, and Developmental Biology, and ³Department of Biochemistry and Biophysics, Yale University, New Haven, Connecticut 06520, USA

Abstract

Functional analysis of the proteome is an essential part of genomic research. To facilitate different proteomic approaches, a MORF (moveable ORF) library of 5854 yeast expression plasmids was constructed, each expressing a sequence-verified ORF as a C-terminal ORF fusion protein, under regulated control. Analysis of 5573 MORFs demonstrates that nearly all verified ORFs are expressed, suggests the authenticity of 48 ORFs characterized as dubious, and implicates specific processes including cytoskeletal organization and transcriptional control in growth inhibition caused by overexpression. Global analysis of glycosylated proteins identifies 109 new confirmed N-linked and 345 candidate glycoproteins, nearly doubling the known yeast glycome.

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Footnotes

Supplemental material is available at http://www.genesdev.org.
Article and publication are at http://www.genesdev.org/cgi/doi/10.1101/gad.1362105.
↵4 These authors contributed equally to this work.
Corresponding authors.
↵5 E-MAIL elizabeth_grayhack{at}urmc.rochester.edu; FAX (585) 271-2683.
↵6 E-MAIL michael.snyder{at}yale.edu; FAX (203) 432-3597.
- Accepted September 26, 2005.
- Received August 8, 2005.
Cold Spring Harbor Laboratory Press