What I wanted to do this summer

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Presentation transcript:

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

Also in SPT3-TAP and SPT7-TAP 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

Also in SPT3-TAP and SPT7-TAP 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