255
protein function in living cells using synthetic
small molecules. Cell 126(5):995–1004
6. Taxis C, Stier G, Spadaccini R, Knop M (2009)
Efficient protein depletion by genetically controlled deprotection of a dormant N-degron.
Mol Syst Biol 5:267
7. Nishimura K, Fukagawa T, Takisawa H,
Kakimoto T, Kanemaki M (2009) An auxinbased degron system for the rapid depletion of
proteins in nonplant cells. Nat Methods
6(12):917–922
8. Jungbluth M, Mosch HU, Taxis C (2012)
Acetate regulation of spore formation is under
the control of the Ras/cyclic AMP/protein
kinase A pathway and carbon dioxide in
Saccharomyces cerevisiae. Eukaryot Cell 11(8):
1021–1032
9. Renicke C, Schuster D, Usherenko S, Essen
LO, Taxis C (2013) A LOV2 domain-based
optogenetic tool to control protein degradation and cellular function. Chem Biol 20(4):
619–626
10. Christie JM, Salomon M, Nozue K, Wada M,
Briggs WR (1999) LOV (light, oxygen, or
voltage) domains of the blue-light photoreceptor phototropin (nph1): binding sites for the
chromophore flavin mononucleotide. Proc
Natl Acad Sci U S A 96(15):8779–8783
11. Harper SM, Neil LC, Gardner KH (2003)
Structural basis of a phototropin light switch.
Science 301(5639):1541–1544
12. Erales J, Coffino P (2014) Ubiquitinindependent
proteasomal
degradation.
Biochim Biophys Acta 1843(1):216–221
13. Takeuchi J, Chen H, Coffino P (2007)
Proteasome substrate degradation requires
association plus extended peptide. EMBO
J 26(1):123–131
14. Takeuchi J, Chen H, Hoyt MA, Coffino P
(2008) Structural elements of the ubiquitinindependent proteasome degron of ornithine
decarboxylase. Biochem J 410(2):401–407
15. Jungbluth M, Renicke C, Taxis C (2010)
Targeted protein depletion in Saccharomyces
cerevisiae by activation of a bidirectional
degron. BMC Syst Biol 4:176
16. Usherenko S, Stibbe H, Musco M, Essen LO,
Kostina EA, Taxis C (2014) Photo-sensitive
degron variants for tuning protein stability by
light. BMC Syst Biol 8:128
17. Pereira G, Tanaka TU, Nasmyth K, Schiebel E
(2001) Modes of spindle pole body inheritance
and segregation of the Bfa1p-Bub2p checkpoint protein complex. EMBO J 20(22):
6359–6370
18. Sherman F (2002) Getting started with yeast.
Methods Enzymol 350:3–41
19. van Leeuwen J, Andrews B, Boone C, Tan G
(2015) Rapid and efficient plasmid construction by homologous recombination in yeast.
Cold Spring Harb Protoc 2015(9): pdb
prot085100
20. Renicke C, Spadaccini R, Taxis C (2013) A
tobacco etch virus protease with increased substrate tolerance at the P1' position. PLoS One
8(6):e67915
21. Ausubel FM, Kingston, R.E., Seidman, F.G.,
Struhl, K., Moore, D.D., Brent, R., and Smith,
F.A. (eds) (1995) Current protocols in molecular biology. John Wiley and Sons, New York,
NY
22. Schiestl RH, Gietz RD (1989) High efficiency
transformation of intact yeast cells using single
stranded nucleic acids as a carrier. Curr Genet
16(5–6):339–346
23. Janke C, Magiera MM, Rathfelder N, Taxis C,
Reber S, Maekawa H, Moreno-Borchart A,
Doenges G, Schwob E, Schiebel E, Knop M
(2004) A versatile toolbox for PCR-based tagging of yeast genes: new fluorescent proteins,
more markers and promoter substitution cassettes. Yeast 21(11):947–962
24. Taxis C, Knop M (2006) System of centromeric, episomal, and integrative vectors based
on drug resistance markers for Saccharomyces
cerevisiae. Biotechniques 40(1):73–78
25. Taxis C, Knop M (2012) TIPI: TEV proteasemediated induction of protein instability.
Methods Mol Biol 832:611–626
26. Lutz AP, Renicke C, Taxis C (2016) Controlling
protein activity and degradation using blue
light. Methods Mol Biol 1408:67–78
27. Yaffe MP, Schatz G (1984) Two nuclear mutations that block mitochondrial protein import
in yeast. Proc Natl Acad Sci U S A 81(15):
4819–4823
28. Laemmli UK (1970) Cleavage of structural
proteins during the assembly of the head of
bacteriophage
T4.
Nature
227(5259):
680–685
29. Towbin H, Staehelin T, Gordon J (1979)
Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci
U S A 76(9):4350–4354
30. Ninnemann H, Butler WL, Epel BL (1970)
Inhibition of respiration in yeast by light.
Biochim Biophys Acta 205(3):499–506
31. Yee EF, Diensthuber RP, Vaidya AT, Borbat PP,
Engelhard C, Freed JH, Bittl R, Moglich A,
Crane BR (2015) Signal transduction in lightoxygen-voltage receptors lacking the adductforming cysteine residue. Nat Commun
6:10079
Controling Protein Stability with Light
protein function in living cells using synthetic
small molecules. Cell 126(5):995–1004
6. Taxis C, Stier G, Spadaccini R, Knop M (2009)
Efficient protein depletion by genetically controlled deprotection of a dormant N-degron.
Mol Syst Biol 5:267
7. Nishimura K, Fukagawa T, Takisawa H,
Kakimoto T, Kanemaki M (2009) An auxinbased degron system for the rapid depletion of
proteins in nonplant cells. Nat Methods
6(12):917–922
8. Jungbluth M, Mosch HU, Taxis C (2012)
Acetate regulation of spore formation is under
the control of the Ras/cyclic AMP/protein
kinase A pathway and carbon dioxide in
Saccharomyces cerevisiae. Eukaryot Cell 11(8):
1021–1032
9. Renicke C, Schuster D, Usherenko S, Essen
LO, Taxis C (2013) A LOV2 domain-based
optogenetic tool to control protein degradation and cellular function. Chem Biol 20(4):
619–626
10. Christie JM, Salomon M, Nozue K, Wada M,
Briggs WR (1999) LOV (light, oxygen, or
voltage) domains of the blue-light photoreceptor phototropin (nph1): binding sites for the
chromophore flavin mononucleotide. Proc
Natl Acad Sci U S A 96(15):8779–8783
11. Harper SM, Neil LC, Gardner KH (2003)
Structural basis of a phototropin light switch.
Science 301(5639):1541–1544
12. Erales J, Coffino P (2014) Ubiquitinindependent
proteasomal
degradation.
Biochim Biophys Acta 1843(1):216–221
13. Takeuchi J, Chen H, Coffino P (2007)
Proteasome substrate degradation requires
association plus extended peptide. EMBO
J 26(1):123–131
14. Takeuchi J, Chen H, Hoyt MA, Coffino P
(2008) Structural elements of the ubiquitinindependent proteasome degron of ornithine
decarboxylase. Biochem J 410(2):401–407
15. Jungbluth M, Renicke C, Taxis C (2010)
Targeted protein depletion in Saccharomyces
cerevisiae by activation of a bidirectional
degron. BMC Syst Biol 4:176
16. Usherenko S, Stibbe H, Musco M, Essen LO,
Kostina EA, Taxis C (2014) Photo-sensitive
degron variants for tuning protein stability by
light. BMC Syst Biol 8:128
17. Pereira G, Tanaka TU, Nasmyth K, Schiebel E
(2001) Modes of spindle pole body inheritance
and segregation of the Bfa1p-Bub2p checkpoint protein complex. EMBO J 20(22):
6359–6370
18. Sherman F (2002) Getting started with yeast.
Methods Enzymol 350:3–41
19. van Leeuwen J, Andrews B, Boone C, Tan G
(2015) Rapid and efficient plasmid construction by homologous recombination in yeast.
Cold Spring Harb Protoc 2015(9): pdb
prot085100
20. Renicke C, Spadaccini R, Taxis C (2013) A
tobacco etch virus protease with increased substrate tolerance at the P1' position. PLoS One
8(6):e67915
21. Ausubel FM, Kingston, R.E., Seidman, F.G.,
Struhl, K., Moore, D.D., Brent, R., and Smith,
F.A. (eds) (1995) Current protocols in molecular biology. John Wiley and Sons, New York,
NY
22. Schiestl RH, Gietz RD (1989) High efficiency
transformation of intact yeast cells using single
stranded nucleic acids as a carrier. Curr Genet
16(5–6):339–346
23. Janke C, Magiera MM, Rathfelder N, Taxis C,
Reber S, Maekawa H, Moreno-Borchart A,
Doenges G, Schwob E, Schiebel E, Knop M
(2004) A versatile toolbox for PCR-based tagging of yeast genes: new fluorescent proteins,
more markers and promoter substitution cassettes. Yeast 21(11):947–962
24. Taxis C, Knop M (2006) System of centromeric, episomal, and integrative vectors based
on drug resistance markers for Saccharomyces
cerevisiae. Biotechniques 40(1):73–78
25. Taxis C, Knop M (2012) TIPI: TEV proteasemediated induction of protein instability.
Methods Mol Biol 832:611–626
26. Lutz AP, Renicke C, Taxis C (2016) Controlling
protein activity and degradation using blue
light. Methods Mol Biol 1408:67–78
27. Yaffe MP, Schatz G (1984) Two nuclear mutations that block mitochondrial protein import
in yeast. Proc Natl Acad Sci U S A 81(15):
4819–4823
28. Laemmli UK (1970) Cleavage of structural
proteins during the assembly of the head of
bacteriophage
T4.
Nature
227(5259):
680–685
29. Towbin H, Staehelin T, Gordon J (1979)
Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci
U S A 76(9):4350–4354
30. Ninnemann H, Butler WL, Epel BL (1970)
Inhibition of respiration in yeast by light.
Biochim Biophys Acta 205(3):499–506
31. Yee EF, Diensthuber RP, Vaidya AT, Borbat PP,
Engelhard C, Freed JH, Bittl R, Moglich A,
Crane BR (2015) Signal transduction in lightoxygen-voltage receptors lacking the adductforming cysteine residue. Nat Commun
6:10079
Controling Protein Stability with Light