Home LiteratureArticle Details
PMID: 10702289 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Functional domain mapping and subcellular distribution of Dal82p in Saccharomyces cerevisiae.

The Journal of biological chemistry ·Vol. 275 ·No. 10 ·2000-03-10 ·Pages 7198-204

Scott S, Dorrington R, Svetlov V, Beeser AE, Distler M, Cooper TG

Abstract

Previous studies have shown that (i) Dal81p and Dal82p are required for allophanate-induced gene expression in Saccharomyces cerevisiae; (ii) the cis-acting element mediating the induced transcriptional response to allophanate is a dodecanucleotide, UIS(ALL); and (iii) Dal82p binds specifically to UIS(ALL). Here we show that Dal82p is localized to the nucleus and parallels movement of the DNA through the cell cycle. Deletion analysis of DAL82 identified and localized three functional domains. Electrophoretic mobility shift assays identified a peptide (consisting of Dal82p amino acids 1-85) that is sufficient to bind a DNA fragment containing UIS(ALL). LexA-tethering experiments demonstrated that Dal82p is capable of mediating transcriptional activation. The activation domain consists of two parts: (i) an absolutely required core region (amino acids 66-99) and (ii) less well defined regions flanking residues 66-99 that are required for full wild-type levels of activation. The Dal82p C terminus contains a predicted coiled-coil motif that down-regulates Dal82p-mediated transcriptional activation.

MeSH Terms
Base Sequence Binding Sites DNA/metabolism Fungal Proteins/analysis,chemistry Membrane Transport Proteins/genetics Molecular Sequence Data Saccharomyces cerevisiae/chemistry Saccharomyces cerevisiae Proteins Transcriptional Activation
Chemicals
DAL4 protein, S cerevisiae DAL5 protein, S cerevisiae Fungal Proteins Membrane Transport Proteins Saccharomyces cerevisiae Proteins DNA
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Scott S
Department of Microbiology and Immunology, University of Tennessee, Memphis, Tennessee 38163, USA.
Dorrington R
Svetlov V
Beeser A E
Distler M
Cooper T G
References (50)
50 references, click to expand
  1. Identification of sequences responsible for transcriptional activation of the allantoate permease gene in Saccharomyces cerevisiae.
    Mol Cell Biol. 1989 Feb;9(2):602-8 PMID: 2651902
  2. The induction of urea carboxylase and allophanate hydrolase in Saccharomyces cerevisiae.
    J Biol Chem. 1973 Sep 10;248(17):6203-9 PMID: 4580053
  3. Upstream induction sequence, the cis-acting element required for response to the allantoin pathway inducer and enhancement of operation of the nitrogen-regulated upstream activation sequence in Saccharomyces cerevisiae.
    J Bacteriol. 1991 Nov;173(22):7186-95 PMID: 1938916
  4. Gzf3p, a fourth GATA factor involved in nitrogen-regulated transcription in Saccharomyces cerevisiae.
    Mol Microbiol. 1997 Mar;23(6):1157-68 PMID: 9106207
  5. Transcription factor-green fluorescent protein chimeric fusion proteins and their use in studies of DNA affinity chromatography.
    J Chromatogr A. 1998 Apr 17;803(1-2):131-9 PMID: 9604329
  6. A gene product needed for induction of allantoin system genes in Saccharomyces cerevisiae but not for their transcriptional activation.
    Mol Cell Biol. 1989 Sep;9(9):3869-77 PMID: 2674683
  7. An adenosine triphosphate-dependent, avidin-sensitive enzymatic cleavage of urea in yeast and green algae.
    J Biol Chem. 1968 Oct 10;243(19):5213-5 PMID: 5679987
  8. Genetic evidence for Gln3p-independent, nitrogen catabolite repression-sensitive gene expression in Saccharomyces cerevisiae.
    J Bacteriol. 1995 Dec;177(23):6910-8 PMID: 7592485
  9. Isolation and characterization of mutants that produce the allantoin-degrading enzymes constitutively in Saccharomyces cerevisiae.
    Mol Cell Biol. 1982 Sep;2(9):1088-95 PMID: 6757722
  10. A family of vectors that facilitate transposon and insertional mutagenesis of cloned genes in yeast.
    Yeast. 1994 Oct;10(10):1267-72 PMID: 7900415
  11. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.
    Genetics. 1989 May;122(1):19-27 PMID: 2659436
  12. Differentially regulated malate synthase genes participate in carbon and nitrogen metabolism of S. cerevisiae.
    Nucleic Acids Res. 1992 Nov 11;20(21):5677-86 PMID: 1454530
  13. Induction and repression of the urea amidolyase gene in Saccharomyces cerevisiae.
    Mol Cell Biol. 1986 Nov;6(11):3954-64 PMID: 3025621
  14. Sequence and expression of GLN3, a positive nitrogen regulatory gene of Saccharomyces cerevisiae encoding a protein with a putative zinc finger DNA-binding domain.
    Mol Cell Biol. 1991 Dec;11(12):6216-28 PMID: 1682800
  15. The Saccharomyces cerevisiae DAL80 repressor protein binds to multiple copies of GATAA-containing sequences (URSGATA).
    J Bacteriol. 1993 Sep;175(18):5851-61 PMID: 8376332
  16. Nitrogen catabolite repression in yeasts and filamentous fungi.
    Adv Microb Physiol. 1985;26:1-88 PMID: 2869649
  17. The green fluorescent protein targets secretory proteins to the yeast vacuole
    Biochim Biophys Acta. 1999 Mar 9;1410(3):287-98 PMID: 10082794
  18. Nuclear localization of the C2H2 zinc finger protein Msn2p is regulated by stress and protein kinase A activity.
    Genes Dev. 1998 Feb 15;12(4):586-97 PMID: 9472026
  19. Transcriptional regulation of the DAL5 gene in Saccharomyces cerevisiae.
    J Bacteriol. 1987 Aug;169(8):3521-4 PMID: 3301804
  20. DAL82, a second gene required for induction of allantoin system gene transcription in Saccharomyces cerevisiae.
    J Bacteriol. 1991 Jan;173(1):255-61 PMID: 1898922
  21. A method for gene disruption that allows repeated use of URA3 selection in the construction of multiply disrupted yeast strains.
    Genetics. 1987 Aug;116(4):541-5 PMID: 3305158
  22. Structure and transcription of the allantoate permease gene (DAL5) from Saccharomyces cerevisiae.
    J Bacteriol. 1988 Jan;170(1):266-71 PMID: 3275614
  23. The DAL7 promoter consists of multiple elements that cooperatively mediate regulation of the gene's expression.
    Mol Cell Biol. 1989 Aug;9(8):3231-43 PMID: 2552287
  24. Complete nucleotide sequence of Saccharomyces cerevisiae chromosome X.
    EMBO J. 1996 May 1;15(9):2031-49 PMID: 8641269
  25. Sequences of two adjacent genes, one (DAL2) encoding allantoicase and another (DCG1) sensitive to nitrogen-catabolite repression in Saccharomyces cerevisiae.
    Gene. 1991 Jul 31;104(1):55-62 PMID: 1916277
  26. Vectors for the inducible overexpression of glutathione S-transferase fusion proteins in yeast.
    Yeast. 1993 Jul;9(7):715-22 PMID: 8368005
  27. Recognition of nitrogen-responsive upstream activation sequences of Saccharomyces cerevisiae by the product of the GLN3 gene.
    J Bacteriol. 1995 Jul;177(14):4190-3 PMID: 7608102
  28. Multifunctional yeast high-copy-number shuttle vectors.
    Gene. 1992 Jan 2;110(1):119-22 PMID: 1544568
  29. Controlling basal expression in an inducible T7 expression system by blocking the target T7 promoter with lac repressor.
    J Mol Biol. 1991 May 5;219(1):45-59 PMID: 1902522
  30. Regulatory circuit for responses of nitrogen catabolic gene expression to the GLN3 and DAL80 proteins and nitrogen catabolite repression in Saccharomyces cerevisiae.
    J Bacteriol. 1993 Jan;175(1):64-73 PMID: 8416910
  31. Positive and negative regulatory elements control the expression of the UGA4 gene coding for the inducible 4-aminobutyric-acid-specific permease in Saccharomyces cerevisiae.
    Eur J Biochem. 1989 May 1;181(2):357-61 PMID: 2653828
  32. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. 1979.
    Biotechnology. 1992;24:145-9 PMID: 1422008
  33. Role of the GATA factors Gln3p and Nil1p of Saccharomyces cerevisiae in the expression of nitrogen-regulated genes.
    Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9450-4 PMID: 7568152
  34. The UGA4 UASNTR site required for GLN3-dependent transcriptional activation also mediates DAL80-responsive regulation and DAL80 protein binding in Saccharomyces cerevisiae.
    J Bacteriol. 1994 Aug;176(15):4718-25 PMID: 8045902
  35. Sequence of the GLN1 gene of Saccharomyces cerevisiae: role of the upstream region in regulation of glutamine synthetase expression.
    J Bacteriol. 1992 Mar;174(6):1828-36 PMID: 1347768
  36. The regulation of urea amidolyase of Saccharomyces cerevisiae: mating type influence on a constitutivity mutation acting in cis.
    Mol Gen Genet. 1978 Nov 9;166(3):251-8 PMID: 368577
  37. What is the function of nitrogen catabolite repression in Saccharomyces cerevisiae?
    J Bacteriol. 1983 Aug;155(2):623-7 PMID: 6135687
  38. Gat1p, a GATA family protein whose production is sensitive to nitrogen catabolite repression, participates in transcriptional activation of nitrogen-catabolic genes in Saccharomyces cerevisiae.
    Mol Cell Biol. 1996 Mar;16(3):847-58 PMID: 8622686
  39. G1n3p is capable of binding to UAS(NTR) elements and activating transcription in Saccharomyces cerevisiae.
    J Bacteriol. 1996 Jun;178(12):3470-9 PMID: 8655543
  40. Requirement of upstream activation sequences for nitrogen catabolite repression of the allantoin system genes in Saccharomyces cerevisiae.
    Mol Cell Biol. 1989 Dec;9(12):5440-4 PMID: 2511434
  41. The minimal transactivation region of Saccharomyces cerevisiae Gln3p is localized to 13 amino acids.
    J Bacteriol. 1997 Dec;179(24):7644-52 PMID: 9401021
  42. The GLN3 gene product is required for transcriptional activation of allantoin system gene expression in Saccharomyces cerevisiae.
    J Bacteriol. 1990 Feb;172(2):1014-8 PMID: 2153652
  43. Pleiotropic control of five eucaryotic genes by multiple regulatory elements.
    J Bacteriol. 1982 Sep;151(3):1237-46 PMID: 7050082
  44. The DAL82 protein of Saccharomyces cerevisiae binds to the DAL upstream induction sequence (UIS).
    Nucleic Acids Res. 1993 Aug 11;21(16):3777-84 PMID: 8367295
  45. The DAL81 gene product is required for induced expression of two differently regulated nitrogen catabolic genes in Saccharomyces cerevisiae.
    Mol Cell Biol. 1991 Feb;11(2):1161-6 PMID: 1990272
  46. Two mutually exclusive regulatory systems inhibit UASGATA, a cluster of 5'-GAT(A/T)A-3' upstream from the UGA4 gene of Saccharomyces cerevisiae.
    Nucleic Acids Res. 1995 Feb 25;23(4):558-64 PMID: 7899075
  47. A cis-acting element present in multiple genes serves as a repressor protein binding site for the yeast CAR1 gene.
    Mol Cell Biol. 1990 Aug;10(8):3884-95 PMID: 2115115
  48. Synergistic operation of the CAR2 (Ornithine transaminase) promoter elements in Saccharomyces cerevisiae.
    J Bacteriol. 1999 Nov;181(22):7052-64 PMID: 10559172
  49. Saturation mutagenesis of the UASNTR (GATAA) responsible for nitrogen catabolite repression-sensitive transcriptional activation of the allantoin pathway genes in Saccharomyces cerevisiae.
    J Bacteriol. 1991 Aug;173(16):4977-82 PMID: 1860815
  50. Cross regulation of four GATA factors that control nitrogen catabolic gene expression in Saccharomyces cerevisiae.
    J Bacteriol. 1997 Jun;179(11):3416-29 PMID: 9171383
Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
0021-9258
DOI
10.1074/jbc.275.10.7198
Published
2000-03-10
Pages
7198-204
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC4384442
Subset
IM
Grants
NIGMS NIH HHS · R01 GM035642 · United States
NIGMS NIH HHS · GM-35642 · United States
External Links
PubMed source DOI: 10.1074/jbc.275.10.7198 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2