This web page was produced as an assignment for an undergraduate course at Davidson College.

 

My Favorite Yeast ORFs: UBI4 and SET4

 

Using Microarrays to Look at Gene Expression

 

 

This web page considers expression of the yeast genes UBI4 and SET4 using data from microarrays.  Information on gene expression from microarrays, or DNA chips, is available from the following databases: Expression Connection, the Yeast Microarray Global Viewer (yMGV), the Stanford Microarray Database (SMD), and the Claude Jacq lab Microarray Page.  Expression of annotated genes involved in known pathways can be seen using the application, GenMAPP.

 

UBI4

UBI4 encodes ubiqutin, a protein found in the cytoplasm of S.cerevisiae involved in protein degradation tagging.  UBI4 expression is associated with stress response.

 

Scale : (fold repression/induction)

ORF

Gene

YLL039C

UBI4

YOR185C

GSP2

YER103W

SSA4

YHR104W

GRE3

Figure 1: Microarray data for UBI4 showing expression in response to stress.  Three similar genes are shown, with a Pearson correlation of > 0.8 to UBI4.  Click on a color strip to see data for that gene.  Courtesy of Expression Connection. 

 

 

According to this data, UBI4 changes its expression over time under conditions of stress.  Expression changes are noticeable for individual stress conditions over time.  UBI4 expression patterns are specific.  It will not be induced in response to any stress, but rather will be induced in situations where the cell needs increased amounts of ubiquitin to target proteins for degradation

 

SET4

SET4 is a non-annotated ORF found on chromosome 10 of S.cerevisiae.  Using the program PREDATOR and Kyte-Doolittle analysis it was predicted that SET4 does not encode a transmembrane protein.  Predictions can be made about SET4’s function by looking at its expression pattern. 

 

A paper by Linde, et al. shows that SET4 is induced under anaerobic conditions.  The cells used in this study are described as follows: “The cultures (strain CEN.PK113-7D (MATa))were fed with a defined mineral medium containing glucose in an Applikon laboratory fermentor. The dilution rate was 0.1h-1 at 30°C, pH was 5.0. Aerobic conditions were maintained by sparging the cultures with air (0.5 liter.min-1). For anaerobic cultures, the medium was supplemented with Tween 80 and ergosterol. Anaerobic conditions were maintained by sparging the medium with pure nitrogen gas (0.5 liter.min-1).” yMGV, 2001

 

Figure 2: This chart shows SET4 induction (red) on a log2 scale for five trials under conditions of anaerbic respiration. The thin red and green lines designate one fold induction and repression.  Experiments were performed by the Linde lab.  Courtesy of yMGV.

 

Only 383 (6.2%) of the 6169 ORFs tested showed 2 fold induction under anaerobic conditions.  SET4 is part of the 6.2%.  SET4’s induction is constant over five trials of the same experiment.  The results are probably not the result of chance.  It can be assumed that SET4 is induced under anaerobic conditions.

 

Using the “guilt by association” method, SET4’s predicted expression pattern could be compared with that of known genes induced under anaerobic respiration.  However, this information is not available.  Instead SET4’s expression pattern will be considered during sporulation.

 

Orf

Gene

Process

Function

Component

YJL105W

SET4

biological_process unknown

molecular_function unknown

not yet annotated

YKL177W

 

biological_process unknown

molecular_function unknown

not yet annotated

YKL058W

TOA2

transcription initiation, from Pol II promoter

general RNA polymerase II transcription factor

transcription factor TFIIA

YGR104C

SRB5

transcription, from Pol II promoter

RNA polymerase II transcription mediator

mediator complex

Figure 3: Microarray data for SET4 showing expression during sporulation.  Three similar genes are shown, with a Pearson correlation of > 0.8 to SET4.  Click on a color strip to see data for that gene.  Courtesy of Expression Connection. 

 

SET4 appears to have similar expression patterns to two known genes, TOA2 and SRB5, during sporulation.  Their gene products are involved in transcription.  Using the “guilt by association” method, it could be assumed that SET4 is also involved in a related pathway.  It should be noted that this relationship is not a result of chromosome location.  SET4, TOA2, and SRB5 are on different chromosomes.

 

SET4 was previously predicted not to be a transmembrane protein.  Using microarray data, it can be predicted that SET4 induces expression under anaerobic conditions and and is involved in transcription with TOA2 and SRB5.


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Updated 10/19/01