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Vol. 12, Issue 7, 2147-2170, July 2001
Department of Molecular, Cellular and Developmental Biology, Yale
University, New Haven, Connecticut 06520-8103
A genome-wide screen of 4168 homozygous diploid yeast deletion
strains has been performed to identify nonessential genes that participate in the bipolar budding pattern. By examining bud scar patterns representing the sites of previous cell divisions, 127 mutants
representing three different phenotypes were found: unipolar, axial-like, and random. From this screen, 11 functional classes of
known genes were identified, including those involved in
actin-cytoskeleton organization, general bud site selection, cell
polarity, vesicular transport, cell wall synthesis, protein
modification, transcription, nuclear function, translation, and other
functions. Four characterized genes that were not known previously to
participate in bud site selection were also found to be important for
the haploid axial budding pattern. In addition to known genes, we found
22 novel genes (20 are designated BUD13-BUD32) important
for bud site selection. Deletion of one resulted in unipolar budding
exclusively from the proximal pole, suggesting that this gene plays an
important role in diploid distal budding. Mutations in 20 other novel
BUD genes produced a random budding phenotype and one
produced an axial-like budding defect. Several of the novel Bud
proteins were fused to green fluorescence protein; two proteins were
found to localize to sites of polarized cell growth (i.e., the bud tip in small budded cells and the neck in cells undergoing cytokinesis), similar to that postulated for the bipolar signals and proteins that
target cell division site tags to their proper location in the cell.
Four others localized to the nucleus, suggesting that they play a role
in gene expression. The bipolar distal marker Bud8 was localized in a
number of mutants; many showed an altered Bud8-green fluorescence
protein localization pattern. Through the genome-wide identification
and analysis of different mutants involved in bipolar bud site
selection, an integrated pathway for this process is presented in which
proximal and distal bud site selection tags are synthesized and
localized at their appropriate poles, thereby directing growth at those
sites. Genome-wide screens of defined collections of mutants hold
significant promise for dissecting many biological processes in yeast.
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