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What I wanted to do this summer
Evaluate inducible inactivation system designed to
regulate mitochondrial gene expression
Examine unintended consequences of TAP-tagging
What I really did this summer
Targets for protein engineering
Pilot studies
= no TAP-tag
FY631
MAT(A) his4-917, lys2-173R2,
leu21, ura3-52, trp163
= SPT3-TAP
= SPT7-TAP
= SPT10-TAP
FY2305
MAT(A) his3200, lys2-128,
leu21, ura3-52, trp163
SPT10: most affected genes, pt 1
higher - TAP
HIS3 as expected
SPT10: most affected genes, pt 1
higher - TAP
LYS2
No TAP
lys2-173R2
vs
+TAP
lys2-128
SPT10: most affected genes, pt 1
higher - TAP
OLI1
again!
F0-ATP synthase
subunit 9 encoded
on the
mitochondrial
genome
Ask:
OligomycinR?
Also in SPT3-TAP
and SPT7-TAP
SPT10: most affected genes, pt 1
higher - TAP
mitochondrial
genes
SPT10: most affected genes, pt 2
higher + TAP
SPT10
Increased expression of
SPT10-TAP vs SPT10
SPT10: most affected genes, pt 2
higher + TAP
PHO5
and
PHO11
SPT10: most affected genes, pt 2
higher + TAP
ADH2 (alcohol
dehydrogenase)
and
ADY2 (Acetate
transporter
required for
normal
sporulation;
phosphorylated
in mitochondria)
Also in SPT3-TAP
and SPT7-TAP
End of summer ppt