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Volume 142, Issue 6

The QUTA activator and QUTR repressor proteins of Aspergillus nidulans interact to regulate transcription of the quinate utilization pathway genes Free

Abstract

Genetic evidence suggests that the activity of the native QUTA transcription activator protein is negated by the action of the QUTR transcription repressor protein. When was transformed with plasmids containing the wild-type gene, transformants that constitutively expressed the quinate pathway enzymes were isolated. The constitutive phenotype of these transformants was associated with an increased copy number of the transforming gene and elevated mRNA levels. Conversely, when was transformed with plasmids containing the gene under the control of the constitutive promoter, transformants with a super-repressed phenotype (unable to utilize quinate as a carbon source) were isolated. The super-repressed phenotype of these transformants was associated with an increased copy number of the transforming gene and elevated mRNA levels. These copy-number-dependent phenotypes argue that the levels of the QUTA and QUTR proteins were elevated in the high-copy-number transformants. When diploid strains were formed by combining haploid strains that contained high copy numbers of either the gene (constitutive phenotype) or the gene (super-repressing; non-inducible phenotype), the resulting diploid phenotype was one of quinate-inducible production of the quinate pathway enzymes, in a manner similar to wild-type. The simplest interpretation of these observations is that the QUTR repressor protein mediates its repressing activity through a direct interaction with the QUTA activator protein. Other possible interpretations are discussed in the text. Experiments in which truncated versions of the QUTA protein were produced in the presence of a wild-type QUTA protein indicate that the QUTR repressor protein recognizes and binds to the C-terminal half of the QUTA activator protein.

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Keyword(s): protein-protein interaction , quinate and transcription regulation
© Society for General Microbiology 1996
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1996-06-01
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References

  1. Anton I.A., Duncan K., Coggins J.R. A eukaryotic repressor protein, the qa-1S gene product of Neurospora crassa, is homologous to part of the arom multifunctional enzyme. J Mol Biol 1987; 197:367–371
     [Google Scholar]
  2. Beri R.K., Whittington H., Roberts C.F., Hawkins A.R. Isolation and characterisation of the positively acting regulatory gene qutA from Aspergillus nidulans. Nucleic Acids Res 1987; 15:7991–8001
     [Google Scholar]
  3. Beri R.K., Grant S., Roberts C.F., Smith M., Hawkins A.R. Selective over-expression of the qutE gene encoding catabolic 3-dehydroquinase in multicopy transformants of Aspergillus nidulans. Biochem J 1990; 265:337–342
     [Google Scholar]
  4. Bugg T.D.H., Alefounder P.R., Abell C. An amino acid sequence motif observed amongst enzymes of the shikimate pathway. Biochem J 1991; 276:841–843
     [Google Scholar]
  5. Buxton F.P., Radford A. Cloning of the structural gene for orotidine 5'-monophosphate carboxylase of Neurospora crassa by expression in Escherichia coli. Mol Gen Genet 1983; 190:403–405
     [Google Scholar]
  6. Case M.E., Giles N.H., Doy C.H. Genetical and biochemical evidence for further interrelationships between the polyaromatic synthetic and the quinate-shikimate catabolic pathways in Neurospora crassa. Genetics 1972; 71:337–348
     [Google Scholar]
  7. Cathala G., Savouret J.F., Mendez B., West B.L., Karin B., Martial J.A., Baxter J.D. A method for isolation of intact translationally active ribonucleic acid. DNA 1983; 2:329–335
     [Google Scholar]
  8. Charles I.G., Keyte J.W., Brammar W.J., Smith M., Hawkins A.R. The isolation and nucleotide sequence of the complex arom locus of Aspergillus nidulans. Nucleic Acids Res 1986; 14:2201–2213
     [Google Scholar]
  9. Clements J.M., Roberts C.F. Molecular cloning of the 3-phosphoglycerate kinase (PGK) gene from Aspergillus nidulans. Curr Genet 1985; 9:293–298
     [Google Scholar]
  10. Clements J.M., Roberts C.F. Transcription and processing signals in the 3-phosphoglycerate kinase (PGK) gene from Aspergillus nidulans. Gene 1986; 14:97–105
     [Google Scholar]
  11. Fidel S., Doonan J.H., Morris N.R. Aspergillus nidulans contains a single actin gene which has unique intron locations and encodes a y-actin gene. Gene 1988; 70:283–293
     [Google Scholar]
  12. Geever R.F., Huiet I., Baum J.A., Tyler B.M., Patel V.B., Rutledge B.J., Case M.E., Giles N.H. DNA sequence, organisation, and regulation of the QA gene cluster of Neurospora crassa. J Mol Biol 1989; 207:15–34
     [Google Scholar]
  13. Giles N.H. The organisation, function and evolution of gene clusters in eukaryotes. Am Nat 1978; 112:641–657
     [Google Scholar]
  14. Giles N.H., Case M.E., Baum J., Geever R., Huiet L., Patel V., Tyler B. Gene organisation and regulation in the qa (quinic acid) gene cluster of Neurospora crassa. Microbiol Rev 1985; 49:338–358
     [Google Scholar]
  15. Glazebrook J.A., Mitchell K., Radford A. Molecular genetic analysis of the pyr-4 gene of Neurospora crassa. Mol Gen Genet 1987; 209:399–402
     [Google Scholar]
  16. Grant S., Roberts C.F., Lamb H.K., Stout M., Hawkins A.R. Genetic regulation of the quinic acid utilization (qut) gene cluster in Aspergillus nidulans. J Gen Microbiol 1988; 134:347–358
     [Google Scholar]
  17. Grewe R., Haendler H. 5-Dehydroquinic acid. Biochem Prep 1966; 11:21–26
     [Google Scholar]
  18. Hawkins A.R. The complex arom locus of Aspergillus nidulans: evidence for multiple gene fusions and convergent evolution. Curr Genet 1987; 11:491–498
     [Google Scholar]
  19. Hawkins A.R., Lamb H.K. The molecular biology of multidomain proteins Selected examples. Eur J Biochem 1995; 232:7–18
     [Google Scholar]
  20. Hawkins A.R., Smith M. Domain structure and interaction within the pentafunctional AROM polypeptide. Eur J Biochem 1991; 196:717–724
     [Google Scholar]
  21. Hawkins A.R., Da Silva A.J., Roberts C.F. Cloning and characterisation of the three enzyme structural genes qutB, qutC and qutE from the quinic acid utilisation gene cluster in Aspergillus nidulans. Curr Genet 1985; 9:305–311
     [Google Scholar]
  22. Hawkins A.R., Lamb H.K., Smith M., Keyte J.W., Roberts C.F. Molecular organisation of the quinic acid utilisation (qut) gene cluster in Aspergillus nidulans. Mol Gen Genet 1988; 214:224–231
     [Google Scholar]
  23. Hawkins A.R., Lamb H.K., Roberts C.F. Structure of the Aspergillus nidulans qut repressor-encoding gene: implications for the regulation of transcription initiation. Gene 1992; 110:109–114
     [Google Scholar]
  24. Hawkins A.R., Lamb H.K., Moore J.D., Charles I.G., Roberts C.F. The pre-chorismate (shikimate) and quinate pathways in filamentous fungi: theoretical and practical aspects. J Gen Microbiol 1993a; 139:2891–2899
     [Google Scholar]
  25. Hawkins A.R., Lamb H.K., Moore J.D., Roberts C.F. Genesis of eukaryotic transcriptional activator and repressor proteins by splitting a multidomain anabolic enzyme. Gene 1993b; 136:49–54
     [Google Scholar]
  26. Hawkins A.R., Moore J.D., Adeokun A.M. Characterisation of the 3-dehydroquinase domain of the pentafunctional AROM protein, and the quinate dehydrogenase from Aspergillus nidulans, and the overproduction of the type II 3-dehydroquinase from Neurospora crassa. Biochem J 1993c; 296:451–157
     [Google Scholar]
  27. Hemsley A., Arnheim N., Toney M.D., Cortopassi G., Galas D.J. A simple method for site-directed mutagenesis using the polymerase chain reaction. Nucleic Acids Res 1989; 17:6545–6551
     [Google Scholar]
  28. Huiet L., Giles N.H. The qa repressor of Neurospora crassa; wild-type and mutant nucleotide sequences. Proc Natl Acad Sci USA 1986; 83:3381–3385
     [Google Scholar]
  29. Johnston M. A model fungal regulatory mechanism: the GAL genes of Saccharomjces cerevisiae. Microbiol Rev 1987; 51:458–476
     [Google Scholar]
  30. Kinghorn J.R., Hawkins A.R. Cloning and expression in Escherichia coli K-12 of the biosynthetic dehyroquinase function of the arom cluster gene from the eukaryote Aspergillus nidulans. Mol Gen Genet 1982; 186:145–152
     [Google Scholar]
  31. Lamb H.K., Hawkins A.R., Smith M., Harvey I.J., Brown J., Turner G., Roberts C.F. Spatial and biological characterisation of the complete quinic acid utilisation gene cluster in Aspergillus nidulans. Mol Gen Genet 1990; 223:17–23
     [Google Scholar]
  32. Lamb H.K., Bagshaw C.R., Hawkins A.R. In vivo overproduction of the pentafunctional AROM polypeptide in Aspergillus nidulans affects metabolic flux in the quinate pathway. Mol Gen Genet 1991; 227:187–196
     [Google Scholar]
  33. Lamb H.K., Van den Hombergh J.P.T.W., Newton G.H., Moore J.D., Roberts C.F., Hawkins A.R. Differential flux through the quinate and shikimate pathways: implications for the channeling hypothesis. Biochem J 1992; 284:181–187
     [Google Scholar]
  34. Lamb H.K., Moore J.D., Lakey J.H., Levett L., Wheeler K.A., Lago H., Coggins J.R., Hawkins A.R. Comparative analysis of the QUTR transcription repressor protein and the three C-terminal domains of the pentafunctional AROM enzyme. Biochem 1996; J313:941–950
     [Google Scholar]
  35. Levesley I., Newton G.H., Lamb H.K., Van Schothorst E., Dalgleish R.W.M., Samson A.C., R.; Roberts C.F., Hawkins A.R. Domain structure and function within the QUTA protein of Aspergillus nidulans: implications for the control of transcription. Microbiology 1996; 142:87–98
     [Google Scholar]
  36. Maniatis T., Fritsch E.F., Sambrook J. Molecular Cloning: a Laboratory Manual 1982 Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  37. Moore J.D., Hawkins A.R. Overproduction of, and interaction within, bifunctional domains from the amino- and carboxy-termini of the pentafunctional AROM protein of Aspergillus nidulans. Mol Gen Genet 1993; 240:92–102
     [Google Scholar]
  38. Moore J.D., Lamb H.K., Garbe T., Servos S., Dougan G., Charles I.G., Hawkins A.R. Inducible overproduction of the Aspergillus nidulans pentafunctional AROM protein and the type-I and -II 3-dehydroquinases from Salmonella typhi and Mycobacterium tuberculosis. Biochem J 1992; 287:173–181
     [Google Scholar]
  39. Nogi Y., f Matsumoto K., Oshima Y. Interaction of super-repressible and dominant constitutive mutations for the synthesis of galactose pathway enzymes in Saccharomyces cerevisiae. Mol Gen Genet 1977; 152:137–144
     [Google Scholar]
  40. Streatfield S.J., Toews S., Roberts C.F. Functional analysis of the expression of the 3'-phosphoglycerate kinase pgk gene in Aspergillus nidulans. Mol Gen Genet 1992; 233:231–240
     [Google Scholar]
  41. Turner G., Ballance D.J. In Proceedings of the Fifth International Symposium on the Genetics of Industrial Micro-Organisms 1987 Edited by Alacevic M., Hraneuli E., Toman Z. Zagreb, Yugoslavia: Pliva;
     [Google Scholar]
  42. Van Den Hombergh J.P.T., W., Moore J.D., Charles I.G., Hawkins A.R. Overproduction in Escherichia coli of the dehydroquinate synthase domain of the Aspergillus nidulans penta-functional AROM protein. Biochem J 1992; 284:861–867
     [Google Scholar]
  43. Walker J.E., Saraste M., Runswick M.J., Gay N.J. Distantly related sequences in the alpha-subunits and beta-subunits of ATP synthases, myosin, kinases, and other ATP-requiring enzymes and a common nucleotide binding fold. EMBO J 1982; 1:945–951
     [Google Scholar]
  44. Wheeler K.A., Lamb H.K., Hawkins A.R. Control of metabolic flux through the quinate pathway in Aspergillus nidulans. Biochem J 1996; 315:195–205
     [Google Scholar]
  45. Whittington H.A., Grant S., Roberts C.F., Lamb H.K., Hawkins A.R. Identification and isolation of a putative permease gene in the quinic acid utilisation (qut) gene cluster of Aspergillus nidulans. Curr Genet 1987; 12:135–139
     [Google Scholar]
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The QUTA activator and QUTR repressor proteins of Aspergillus nidulans interact to regulate transcription of the quinate utilization pathway genes
Microbiology 142, 1477 (1996); https://doi.org/10.1099/13500872-142-6-1477
/content/journal/micro/10.1099/13500872-142-6-1477
/content/journal/micro/10.1099/13500872-142-6-1477
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