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

Research output: Contribution to journalArticlepeer-review

386 Scopus citations

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.

Original languageEnglish
Pages (from-to)2816-2826
Number of pages11
JournalGenes and Development
Volume19
Issue number23
DOIs
StatePublished - Dec 1 2005

Keywords

  • Biochemical genomics
  • Galactose lethality
  • Glycosylation
  • High-throughput expression
  • Protein microarray
  • Proteome

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