Analysis of peroxisome assembly and function by combining large-scale
genomic and proteomic analyses.
Marcello Marelli (1), Jennifer J. Smith (1), Rowan H. Christmas
(1), Eugene Yi (1), David Goodlett (1), Richard A. Rachubinski (2), John
D. Aitchison (1)
(1) Laboratory of Yeast Biology, Institute for Systems Biology, 1441
North 34th St, Seattle, WA 98103-8904, USA;
(2) Department of Cell Biology, 5-14 Medical Sciences Building,
University of Alberta, Edmonton, Alberta, T6G 2H7, Canada
We have combined comprehensive proteomic and genomic approaches to
define proteins involved in peroxisome function and biogenesis. The
changes in the yeast transcriptome at various time points during
peroxisome induction and repression were determined by whole-genome
microarray analysis. Pattern-matching the expression profiles using
clustering algorithms identified genes with profiles similar to those of
genes involved in peroxisome development or function. In addition, two
novel proteomics procedures were used to identify the complete protein
complement of yeast peroxisomes. The first of these, applied to purified
peroxisomes, is a highly sensitive gas-phase procedure designed to
identify all proteins in a complex mixture of purified proteins. This
was coupled with a second procedure to identify proteins that
specifically enrich with peroxisomes, thereby eliminating contaminants.
Each of these studies has generated a list of genes that are implicated
in peroxisome function and/or biogenesis including those that encode
known peroxisomal proteins, and known peroxins (proteins required for
peroxisome assembly or maintenance). In addition, several previously
uncharacterized genes were identified including YPL112C, and
YOR084W, which encode proteins of the peroxisomal membrane and
matrix respectively. Ypl112p (Pex25p) encodes a novel peroxin required
for the regulation of peroxisome size.
Return to YGM 2002 Home at SGD