identification of 23 complementation groups required of ... · identification of 23 complementation...
TRANSCRIPT
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Identification of 23 Identification of 23 Complementation Groups Complementation Groups
Required of PostRequired of Post--translational Events in the translational Events in the Yeast Secretory PathwayYeast Secretory Pathway
Peter Novick, Charles Field, Randy SchekmanCell, 21:205-215
Present by Liang
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BackgroundBackground
•Goal:study secretory process
•Basic:complementation test
http://saturn.roswellpark.org/cmb/huberman/BIR572/Fig7YeastGenetics.html
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Research StrategyResearch Strategy
1. Mutation
2. Selection
3. Comp. Test
4. Identify
5. Discussion
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Density EnrichmentDensity Enrichment
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Complementation TestComplementation Test
Total: 72 98% 116 100%
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25C, 25C, InvertaseInvertase AccumulationAccumulation
> >
< >
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25C, Acid 25C, Acid PhosphatasePhosphatase & Sulfate & Sulfate PermeasePermease DefectDefect
= =
< >
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TSEM of Cells, 25C v.s. 37CTSEM of Cells, 25C v.s. 37C
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Organelles AccumulationOrganelles Accumulation
Accumulated Structures:Accumulated Structures:
• Vesicle• Berkeley Body• ER• Small Vesicle
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nuclear poreER
nuclear membrane
plasma membrane
small vesicle
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Berkeley Body
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Berkeley Body
ER
vesicle
vesicle
Berkeley Body
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ConclusionConclusion
•Ludox Density Gradient accumulated secretion mutants.
•At least 23 genes associate with yeast secretory pathway.
•Membrane-enclosed organelles take part in the secretory pathway:ER => ER => BbsBbs => vesicles => cell surface=> vesicles => cell surface
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DisscussionDisscussion
•Eliminate some mutants•TSEM artifact•One comp. group <> one gene
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AppendixAppendix• Yeast Introduction. Cells of the alpha mating type are grown overnight
on agar medium, a high concentration of the pheromone accumulates in the agar surrounding the growth. Then if cells of the a mating type are placed on this agar, they begin to undergo the "shmoo" transformation within a couple of hours.
people.uis.edu/rmosh1/Yeastintromaterial02.pdf