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Gal4 transcription factor

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Regulatory protein GAL4
Identifiers
OrganismSaccharomyces cerevisiae
SymbolGAL4
Entrez855828
UniProtP04386
Search for
StructuresSwiss-model
DomainsInterPro

TheGal4 transcription factoris a positive regulator of gene expression of galactose-induced genes.[1]This protein represents a large fungal family of transcription factors, Gal4 family, which includes over 50 members in the yeastSaccharomyces cerevisiaee.g. Oaf1, Pip2, Pdr1, Pdr3, Leu3.[2]

Gal4 recognizes genes with UASG,anupstream activating sequence,and activates them. In yeast cells, the principal targets are GAL1 (galactokinase), GAL10 (UDP-glucose 4-epimerase), and GAL7 (galactose-1-phosphate uridylyltransferase), three enzymes required for galactose metabolism. This binding has also proven useful in constructing theGAL4/UAS system,a technique for controlling expression in insects.[3]In yeast, Gal4 is by default repressed by Gal80, and activated in the presence ofgalactoseas Gal3 binds away Gal80.[4]

Domains

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Two executive domains, DNA binding and activation domains, provide key function of the Gal4 protein conforming to most of the transcription factors.

Gal4 domains and regulation

DNA binding

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Gal4 N-terminus is a zinc finger and belongs to theZn(2)-C6fungal family. It forms a Zn – cysteines thiolate cluster,[5][6]and specifically recognizes UASGin GAL1 promoter.[7][8]

Gal4 transactivation

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Localised to the C-terminus, belongs to the nine amino acidstransactivation domainfamily, 9aaTAD, together with Oaf1, Pip2, Pdr1, Pdr3, but alsop53,E2A,MLL.[9][10]

Regulation

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Galactoseinduces Gal4 mediated transcription albeit Glucose causes severe repression.[11][12]

As a part of the Gal4 regulation, inhibitory protein Gal80 recognises and binds to the Gal4 region (853-874 aa).[13][14][15]

The inhibitory protein Gal80 is sequestered by regulatory protein Gal3 in Galactose dependent manner. This allows for Gal4 to work when there is galactose.[16][4][17][18]

Mutants

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The Gal4 loss-of-function mutant gal4-64 (1-852 aa, deletion of the Gal4 C-terminal 29 aa) lost both interaction with Gal80 and activation function.[19][20][21]

In the Gal4 reverted mutant Gal4C-62 mutant,[22]a sequence (QTAY N AFMN) with the 9aaTAD pattern emerged and restored activation function of the Gal4 protein.

Inactive constructs

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The activation domain Gal4 is inhibited by C-terminal domain in some Gal4 constructs.[23][24]

Function

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Target

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Transcription

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The Gal4 activation function is mediated byMED15(Gal11).[25][26][27][28][29][30][31]

The Gal4 protein interacts also with other mediators of transcription as are Tra1,[32][33][34]TAF9,[35]and SAGA/MED15 complex.[36][37]

Proteosome

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A subunit of the 26 S proteasome Sug2 regulatory protein has a molecular and functional interaction with Gal4 function.[38][39]Proteolytic turnover of the Gal4 transcription factor is not required for function in vivo.[40]The native Gal4 monoubiquitination protects from 19S-mediated destabilizing under inducing conditions.[41]

Application

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The broad use of the Gal4 is in yeasttwo-hybrid screeningto screen or to assay protein-protein interactions in eukaryotic cells from yeast to human.

In theGAL4/UAS system,the Gal4 protein and Gal4 upstream activating region (UAS) are used to study the gene expression and function in organisms such as the fruit fly.[3]

The Gal4 and inhibitory protein Gal80 have found application in a genetics technique for creating individually labeled homozygous cells calledMARCM(Mosaic analysis with a repressible cell marker).

See also

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References

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  1. ^Klar AJ, Halvorson HO (1974). "Studies on the positive regulatory gene, GAL4, in regulation of galactose catabolic enzymes in Saccharomyces cerevisiae".Molecular & General Genetics.135(3): 203–12.doi:10.1007/BF00268616.PMID4376212.S2CID26014344.
  2. ^Schjerling P, Holmberg S (December 1996)."Comparative amino acid sequence analysis of the C6 zinc cluster family of transcriptional regulators".Nucleic Acids Research.24(23): 4599–607.doi:10.1093/nar/24.23.4599.PMC146297.PMID8967907.
  3. ^abDuffy JB (2002)."GAL4 system in Drosophila: a fly geneticist's Swiss army knife".Genesis.34(1–2): 1–15.doi:10.1002/gene.10150.PMID12324939.S2CID5073328.
  4. ^abJiang F, Frey BR, Evans ML, Friel JC, Hopper JE (October 2009)."Gene activation by dissociation of an inhibitor from a transcriptional activation domain".Molecular and Cellular Biology.29(20): 5604–10.doi:10.1128/MCB.00632-09.PMC2756894.PMID19651897.
  5. ^Marmorstein R, Carey M, Ptashne M, Harrison SC (April 1992). "DNA recognition by GAL4: structure of a protein-DNA complex".Nature.356(6368): 408–14.Bibcode:1992Natur.356..408M.doi:10.1038/356408a0.PMID1557122.S2CID4344434.
  6. ^Pan T, Coleman JE (March 1990). "The DNA binding domain of GAL4 forms a binuclear metal ion complex".Biochemistry.29(12): 2023–9.doi:10.1021/bi00464a019.PMID2186803.
  7. ^Keegan L, Gill G, Ptashne M (February 1986). "Separation of DNA binding from the transcription-activating function of a eukaryotic regulatory protein".Science.231(4739): 699–704.Bibcode:1986Sci...231..699K.doi:10.1126/science.3080805.PMID3080805.
  8. ^Giniger E, Varnum SM, Ptashne M (April 1985). "Specific DNA binding of GAL4, a positive regulatory protein of yeast".Cell.40(4): 767–74.doi:10.1016/0092-8674(85)90336-8.PMID3886158.S2CID31663066.
  9. ^Ding WV, Johnston SA (May 1997)."The DNA binding and activation domains of Gal4p are sufficient for conveying its regulatory signals".Molecular and Cellular Biology.17(5): 2538–49.doi:10.1128/MCB.17.5.2538.PMC232103.PMID9111323.
  10. ^Melcher K, Johnston SA (May 1995)."GAL4 interacts with TATA-binding protein and coactivators".Molecular and Cellular Biology.15(5): 2839–48.doi:10.1128/MCB.15.5.2839.PMC230515.PMID7739564.
  11. ^Klar AJ, Halvorson HO (1974). "Studies on the positive regulatory gene, GAL4, in regulation of galactose catabolic enzymes in Saccharomyces cerevisiae".Molecular & General Genetics.135(3): 203–12.doi:10.1007/BF00268616.PMID4376212.S2CID26014344.
  12. ^Griggs DW, Johnston M (October 1991)."Regulated expression of the GAL4 activator gene in yeast provides a sensitive genetic switch for glucose repression".Proceedings of the National Academy of Sciences of the United States of America.88(19): 8597–601.Bibcode:1991PNAS...88.8597G.doi:10.1073/pnas.88.19.8597.PMC52556.PMID1924319.
  13. ^Kumar PR, Yu Y, Sternglanz R, Johnston SA, Joshua-Tor L (February 2008)."NADP regulates the yeast GAL induction system".Science.319(5866): 1090–2.Bibcode:2008Sci...319.1090K.doi:10.1126/science.1151903.PMC2726985.PMID18292341.
  14. ^Thoden JB, Ryan LA, Reece RJ, Holden HM (October 2008)."The interaction between an acidic transcriptional activator and its inhibitor. The molecular basis of Gal4p recognition by Gal80p".The Journal of Biological Chemistry.283(44): 30266–72.doi:10.1074/jbc.M805200200.PMC2573081.PMID18701455.
  15. ^Johnston SA, Salmeron JM, Dincher SS (July 1987). "Interaction of positive and negative regulatory proteins in the galactose regulon of yeast".Cell.50(1): 143–6.doi:10.1016/0092-8674(87)90671-4.PMID3297350.S2CID46090047.
  16. ^Egriboz O, Jiang F, Hopper JE (November 2011)."Rapid GAL gene switch of Saccharomyces cerevisiae depends on nuclear Gal3, not nucleocytoplasmic trafficking of Gal3 and Gal80".Genetics.189(3): 825–36.doi:10.1534/genetics.111.131839.PMC3213366.PMID21890741.
  17. ^Peng G, Hopper JE (June 2002)."Gene activation by interaction of an inhibitor with a cytoplasmic signaling protein".Proceedings of the National Academy of Sciences of the United States of America.99(13): 8548–53.Bibcode:2002PNAS...99.8548P.doi:10.1073/pnas.142100099.PMC124307.PMID12084916.
  18. ^Tsuyumu S, Adams BG (July 1974)."Dilution kinetic studies of yeast populations: in vivo aggregation of galactose utilizing enzymes and positive regulator molecules".Genetics.77(3): 491–505.doi:10.1093/genetics/77.3.491.PMC1213142.PMID4369925.
  19. ^Douglas HC, Condie F (December 1954)."The genetic control of galactose utilization in Saccharomyces".Journal of Bacteriology.68(6): 662–70.doi:10.1128/jb.68.6.662-670.1954.PMC386212.PMID13221541.
  20. ^Douglas HC, Hawthorne DC (May 1964)."Enzymatic Expression and Genetic Linkage of Genes Controlling Galactose Utilization in Saccharomyces".Genetics.49(5): 837–44.doi:10.1093/genetics/49.5.837.PMC1210618.PMID14158615.
  21. ^Matsumoto K, Adachi Y, Toh-e A, Oshima Y (February 1980)."Function of positive regulatory gene gal4 in the synthesis of galactose pathway enzymes in Saccharomyces cerevisiae: evidence that the GAL81 region codes for part of the gal4 protein".Journal of Bacteriology.141(2): 508–27.doi:10.1128/JB.141.2.508-527.1980.PMC293654.PMID6988385.
  22. ^Johnston SA, Salmeron JM, Dincher SS (July 1987). "Interaction of positive and negative regulatory proteins in the galactose regulon of yeast".Cell.50(1): 143–6.doi:10.1016/0092-8674(87)90671-4.PMID3297350.S2CID46090047.
  23. ^Ma J, Ptashne M (March 1987). "Deletion analysis of GAL4 defines two transcriptional activating segments".Cell.48(5): 847–53.doi:10.1016/0092-8674(87)90081-X.PMID3028647.S2CID4979320.
  24. ^Warfield L, Tuttle LM, Pacheco D, Klevit RE, Hahn S (August 2014)."A sequence-specific transcription activator motif and powerful synthetic variants that bind Mediator using a fuzzy protein interface".Proceedings of the National Academy of Sciences of the United States of America.111(34): E3506-13.Bibcode:2014PNAS..111E3506W.doi:10.1073/pnas.1412088111.PMC4151740.PMID25122681.
  25. ^Fassler JS, Winston F (December 1989)."The Saccharomyces cerevisiae SPT13/GAL11 gene has both positive and negative regulatory roles in transcription".Molecular and Cellular Biology.9(12): 5602–9.doi:10.1128/MCB.9.12.5602.PMC363730.PMID2685570.
  26. ^Han Y, Kodadek T (May 2000)."Peptides selected to bind the Gal80 repressor are potent transcriptional activation domains in yeast".The Journal of Biological Chemistry.275(20): 14979–84.doi:10.1074/jbc.275.20.14979.PMID10809742.
  27. ^Hashimoto H, Kikuchi Y, Nogi Y, Fukasawa T (1983). "Regulation of expression of the galactose gene cluster in Saccharomyces cerevisiae. Isolation and characterization of the regulatory gene GAL4".Molecular & General Genetics.191(1): 31–8.doi:10.1007/BF00330886.PMID6350827.S2CID115216273.
  28. ^Long RM, Mylin LM, Hopper JE (April 1991)."GAL11 (SPT13), a transcriptional regulator of diverse yeast genes, affects the phosphorylation state of GAL4, a highly specific transcriptional activator".Molecular and Cellular Biology.11(4): 2311–4.doi:10.1128/MCB.11.4.2311.PMC359938.PMID2005915.
  29. ^Nogi Y, Fukasawa T (October 1980). "A novel mutation that affects utilization of galactose in Saccharomyces cerevisiae".Current Genetics.2(2): 115–20.doi:10.1007/BF00420623.PMID24189802.S2CID12635991.
  30. ^Sakurai H, Hiraoka Y, Fukasawa T (September 1993)."Yeast GAL11 protein is a distinctive type transcription factor that enhances basal transcription in vitro".Proceedings of the National Academy of Sciences of the United States of America.90(18): 8382–6.Bibcode:1993PNAS...90.8382S.doi:10.1073/pnas.90.18.8382.PMC47360.PMID8378310.
  31. ^Suzuki Y, Nogi Y, Abe A, Fukasawa T (October 1992)."GAL11 protein, an auxiliary transcription activator for genes encoding galactose-metabolizing enzymes in Saccharomyces cerevisiae".Molecular and Cellular Biology.12(10): 4806.doi:10.1128/MCB.12.10.4806.PMC360407.PMID1406662.
  32. ^Lin L, Chamberlain L, Zhu LJ, Green MR (February 2012)."Analysis of Gal4-directed transcription activation using Tra1 mutants selectively defective for interaction with Gal4".Proceedings of the National Academy of Sciences of the United States of America.109(6): 1997–2002.Bibcode:2012PNAS..109.1997L.doi:10.1073/pnas.1116340109.PMC3277556.PMID22308403.
  33. ^Majmudar CY, Labut AE, Mapp AK (July 2009)."Tra1 as a screening target for transcriptional activation domain discovery".Bioorganic & Medicinal Chemistry Letters.19(14): 3733–5.doi:10.1016/j.bmcl.2009.05.045.PMC4322765.PMID19497740.
  34. ^Reeves WM, Hahn S (October 2005)."Targets of the Gal4 transcription activator in functional transcription complexes".Molecular and Cellular Biology.25(20): 9092–102.doi:10.1128/MCB.25.20.9092-9102.2005.PMC1265783.PMID16199885.
  35. ^Klein J, Nolden M, Sanders SL, Kirchner J, Weil PA, Melcher K (February 2003)."Use of a genetically introduced cross-linker to identify interaction sites of acidic activators within native transcription factor IID and SAGA".The Journal of Biological Chemistry.278(9): 6779–86.doi:10.1074/jbc.M212514200.PMID12501245.
  36. ^Larschan E, Winston F (January 2005)."The Saccharomyces cerevisiae Srb8-Srb11 complex functions with the SAGA complex during Gal4-activated transcription".Molecular and Cellular Biology.25(1): 114–23.doi:10.1128/MCB.25.1.114-123.2005.PMC538787.PMID15601835.(http://mcb.asm.org/content/25/1/114/F8.large.jpg)
  37. ^Larsson M, Uvell H, Sandström J, Rydén P, Selth LA, Björklund S (2013)."Functional studies of the yeast med5, med15 and med16 mediator tail subunits".PLOS ONE.8(8): e73137.Bibcode:2013PLoSO...873137L.doi:10.1371/journal.pone.0073137.PMC3750046.PMID23991176.
  38. ^Chang C, Gonzalez F, Rothermel B, Sun L, Johnston SA, Kodadek T (August 2001)."The Gal4 activation domain binds Sug2 protein, a proteasome component, in vivo and in vitro".The Journal of Biological Chemistry.276(33): 30956–63.doi:10.1074/jbc.M102254200.PMID11418596.
  39. ^Russell SJ, Johnston SA (March 2001)."Evidence that proteolysis of Gal4 cannot explain the transcriptional effects of proteasome ATPase mutations".The Journal of Biological Chemistry.276(13): 9825–31.doi:10.1074/jbc.M010889200.PMID11152478.
  40. ^Nalley K, Johnston SA, Kodadek T (August 2006). "Proteolytic turnover of the Gal4 transcription factor is not required for function in vivo".Nature.442(7106): 1054–7.Bibcode:2006Natur.442.1054N.doi:10.1038/nature05067.PMID16929306.S2CID1926315.
  41. ^Ferdous A, Sikder D, Gillette T, Nalley K, Kodadek T, Johnston SA (January 2007)."The role of the proteasomal ATPases and activator monoubiquitylation in regulating Gal4 binding to promoters".Genes & Development.21(1): 112–23.doi:10.1101/gad.1493207.PMC1759896.PMID17167105.

Further reading

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