Transcript UBA4 - SPUR - University of Oregon

The role of UBA4 in the genome of
the yeast Saccharomyces cerevisiae
http://www.chateauneuf.dk/artikler/vini15.jpg
David Roberts, Claire Romelfanger, George Sprague PhD,
Morehouse College, Atlanta GA, University of Oregon, Eugene
OR
Saccharomyces cerevisiae
http://www.chateauneuf.dk/artikler/vi
ni15.jpg
http://www.science.lsus.edu/docu
ments/resume/94859/yeast3.jpg
-Easy to culture
-Eukaryotic cells similar in complexity to plant and animal cells
-Reproduce sexually and asexually
-Exists in haploid and diploid states
-70% of the genome well elucidated
UBA4
Known functions of UBA4
UBA4 is a critical enzyme in the process of urmylation. Urmylation is involved in the
following processes
-Urm1 activating enzyme (urmylation)
-Cell budding
-Invasive growth
-Psuedohyphal growth
-Response to oxidative stress
Why study UBA4?
-UBA4 is present in all eukaryotic organisms
-To establish the specific role of urmylation in its processes
-To establish UBA4 functioning beyond urmylation
Objective of study:
-To gain further insight into the genetic interactions of UBA4
Ubiquitination and Urmylation
Ubi4
Urm1
E1
Uba1Activating
EnzymeUba4
E2
Ubc4Conjugating Enzyme ?
E3
Protein Ligase
Ubr2
?
Ubiquitin Protease
UBP2
?
Synthetic Genetic Analysis
UBA4
Approximately 5000 single
gene deleted mutants
uba∆
YFG
yfg∆
Healthy
Synthetic
Sick
Synthetic
Lethal
Results
Synthetic sick mutant
Plate of mutants
Synthetic lethal mutant
Synthetic Sickness and Lethality
Mutants said to be synthetically sick demonstrated diminished growth when
compared to healthy growing (wild type) mutants.
Synthetically lethal mutants demonstrated no growth when compared to wild type
mutants.
UBA4 synthetic lethal/sick genes sorted by function
Signal Transduction
Cell Organization
and Biogenesis
Protein Degredation
Metabolism
Transcription
RNA Processing
Transport
Unknown
DNA Repair
Protein Transport
DNA Metabolism
UBA4 synthetic lethal/sick genes that overlap with URM1
synthetic lethal/sick genes
Cell organization and
biogenesis
Protein Transport
Unknown
Signal Transduction
Metabolism
Protein synthesis
Conclusions
The results suggest that UBA4 genetically interacts with 56 different genes which participate in the following
cellular processes:
-transport
-cell organization and biogenesis
-transcription
-RNA processing
-DNA repair
-DNA metabolism
-protein transport
-protein degradation
-signal transduction
-metabolism
Furthermore 12 of these genes are known to be synthetically lethal or sick with urm1∆, suggesting that
urmylation may be involved in the processes governed by these genes.
Projections for Further Study
-Subsequent synthetic genetic analysis trials must be conducted to determine
whether or not the results obtained in this experiment are reproducable.
-Random spore analysis must be conducted to verify the existence of the genetic
interactions believed to have been observed.
Acknowledgements
Spur Program
Students
Coordinators
Sprague Lab
Claire Romelfanger
Charles Kubicek
Kimberly Landreth
Steven Tersigni
Dr. George Sprague