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Regulation of lipid storage in Saccharomyces cerevisiae Shaelie Harper McDonough-Stukey Lab A Little History • How do cells “know” when they have made enough lipid? • Previously, Cheryl Jacobs had isolated a mutant (114) we believe is defective at sensing lipid amounts • My work this summer was to begin characterization of the mutant Growth Phenotype (clumpy vs. normal) • Within a tetrad, it can vary what strains are clumpy and which aren’t • If the ratio is 2:2 within a tetrad, this means there is only 1 mutation Clumpy Normal Gas Chromatography data •To the top is a picture of the GC at Hope College •To the left is an example of the data collected by the GC •Each Peak represents a different fatty acid, and its relative amount present in each cell Data Tables •By creating data tables, it is easier to make comparisons and conclusions for each individual strain Fatty acid Amount and Composition Whole cell FA composition Comparison of whole cell FA quantity FA amount (normalized total GC peak area) 50 Percent of total FA sup114 40 diploid w303-1a 30 20 10 0 10:0 12:0 14:0 16:0 16:1 FA species 18:0 18:1 26:0 16000 14000 12000 10000 8000 6000 4000 2000 0 1A 1B 1C 1D Strain •These charts are compiled after analyzing the data from the GC and organizing it onto spreadsheets (shown on the previous slide), so we can compare different strains Result summary Tetrad 1 2 3 4 6 7 8 Spore FA Amount Clumpy Growth A Wt NC B Wt NC C +++ C D +++ C A NC B C C C D NC A C B Wt NC C +++ C D NC A NC B C C NC D C A NC B C C C D NC A NC B C C C D NC A Wt NC B +++ C C +++ C D wt NC •Clumpy Growth is inherited in a single gene manner (2:2 inheritance) •Preliminary results indicate that elevated fatty acid amount is correlated to growth habit, and is a single gene mutation Conclusions • By examining inheritance patterns we are able to see – Clumpy growth habit is correlated to elevated FA amount – Elevated FA amount is a recessive single gene trait Credits • REACH • Dr. McDonough and Dr. Stukey • Rachel Van Kempen • Caitlin Rice