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A new genetic method for isolating functionally interacting genes: high plo1⁺-dependent mutants and their suppressors define genes in mitotic and septation pathways in fission yeast

Cullen, C. Fiona and May, Karen M. and Hagan, Iain M. and Glover, David M. and Ohkura, Hiroyuki (2000) A new genetic method for isolating functionally interacting genes: high plo1⁺-dependent mutants and their suppressors define genes in mitotic and septation pathways in fission yeast. Genetics, 155 (4). pp. 1521-1534. ISSN 0016-6731. PMCID PMC1461180. https://resolver.caltech.edu/CaltechAUTHORS:20201005-153558889

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Abstract

We describe a general genetic method to identify genes encoding proteins that functionally interact with and/or are good candidates for downstream targets of a particular gene product. The screen identifies mutants whose growth depends on high levels of expression of that gene. We apply this to the plo1⁺ gene that encodes a fission yeast homologue of the polo-like kinases. plo1⁺ regulates both spindle formation and septation. We have isolated 17 high plo1⁺-dependent (pld) mutants that show defects in mitosis or septation. Three mutants show a mitotic arrest phenotype. Among the 14 pld mutants with septation defects, 12 mapped to known loci: cdc7, cdc15, cdc11 spg1, and sid2. One of the pld mutants, cdc7-PD1, was selected for suppressor analysis. As multicopy suppressors, we isolated four known genes involved in septation in fission yeast: spg1⁺, sce3⁺, cdc8⁺, and rho1⁺, and two previously uncharacterized genes, mpd1⁺ and mpd2⁺. mpd1⁺ exhibits high homology to phosphatidylinositol 4-phosphate 5-kinase, while mpd2⁺ resembles Saccharomyces cerevisiae SMY2; both proteins are involved in the regulation of actin-mediated processes. As chromosomal suppressors of cdc7-PD1, we isolated mutations of cdc16 that resulted in multiseptation without nuclear division. cdc16⁺, dma1⁺, byr3⁺, byr4⁺ and a truncated form of the cdc7 gene were isolated by complementation of one of these cdc16 mutations. These results demonstrate that screening for high dose-dependent mutants and their suppressors is an effective approach to identify functionally interacting genes.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://www.genetics.org/content/155/4/1521.longPublisherArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1461180/PubMed CentralArticle
ORCID:
AuthorORCID
Glover, David M.0000-0003-0956-0103
Additional Information:© 2000 by the Genetics Society of America. Received February 2, 2000. Accepted April 10, 2000. We thank Drs. A. Carr, K. Maundrell, P. Nurse, and K. Gould for a genomic library, expression vectors, cdc mutants, and sid mutants. This work is supported by the Wellcome Trust and the Cancer Research Campaign.
Funders:
Funding AgencyGrant Number
Wellcome TrustUNSPECIFIED
Cancer Research CampaignUNSPECIFIED
Issue or Number:4
PubMed Central ID:PMC1461180
Record Number:CaltechAUTHORS:20201005-153558889
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20201005-153558889
Official Citation:A New Genetic Method for Isolating Functionally Interacting Genes: High plo1+-Dependent Mutants and Their Suppressors Define Genes in Mitotic and Septation Pathways in Fission Yeast C. Fiona Cullen, Karen M. May, Iain M. Hagan, David M. Glover and Hiroyuki Ohkura GENETICS August 1, 2000 vol. 155 no. 4 1521-1534
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:105822
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:06 Oct 2020 14:12
Last Modified:06 Oct 2020 14:12

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