Abstract
The Dictyostelium discoidin genes are induced in bacteria-grown cells shortly before the onset of development but are also highly expressed during growth in axenic medium. We here show that axenically growing cells strongly respond to the extracellular signal folate by suppressing discoidin synthesis while cell growth and development is not substantially affected. Repression occurs via two previously identified promoter elements, the dIE and the dAXE. Removal of the signal molecules or setting cells up for development results in rapid reactivation of the promoter. Based on this observation, we constructed the transformation vector pVEII and describe a convenient method which allows for controlled expression of a gene of interest in growing cells and also for external modulation in early development. Deletion constructs of the discoidin promoter can be used in addition to vary transcriptional activity over about one order of magnitude.
MeSH Terms
Animals
Base Sequence
DNA, Single-Stranded
Dictyostelium/drug effects,genetics,growth & development
Discoidins
Folic Acid/pharmacology
Fungal Proteins/genetics
Gene Expression Regulation/drug effects
Lectins
Molecular Sequence Data
Promoter Regions, Genetic
Protozoan Proteins
Restriction Mapping
Transcription, Genetic/drug effects
Transformation, Genetic
Chemicals
DNA, Single-Stranded
Discoidins
Fungal Proteins
Lectins
Protozoan Proteins
Folic Acid
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Blusch J
Max-Planck-Institut für Biochemie, Abt. Zellbiologie, Martinsried, Germany.
Morandini P
Nellen W
References (21)
21 references, click to expand
-
Regulation of the Discoidin I gamma gene in Dictyostelium discoideum: identification of individual promoter elements mediating induction of transcription and repression by cyclic AMP.
Mol Cell Biol. 1990 Aug;10(8):4080-8
PMID: 2196444
-
The Dictyostelium discoideum EB4 gene product and a truncated mutant form of the protein are localized in prespore vesicles but absent from mature spores.
Dev Biol. 1991 Mar;144(1):212-4
PMID: 1995398
-
A colony-blot technique for the detection of specific transcripts in eukaryotes.
Anal Biochem. 1989 Jan;176(1):78-81
PMID: 2653101
-
Homologous recombination in the Dictyostelium alpha-actinin gene leads to an altered mRNA and lack of the protein.
EMBO J. 1987 Dec 20;6(13):4143-8
PMID: 2832153
-
Analysis of the prestarvation response in growing cells of Dictyostelium discoideum.
Dev Genet. 1988;9(4-5):315-26
PMID: 3072133
-
Density-dependent induction of discoidin-I synthesis in exponentially growing cells of Dictyostelium discoideum.
Differentiation. 1987;34(2):79-87
PMID: 3622952
-
Phenocopy of discoidin I-minus mutants by antisense transformation in Dictyostelium.
Cell. 1985 Dec;43(3 Pt 2):633-41
PMID: 4075402
-
High-copy-number transformants and co-transformation in Dictyostelium.
Gene. 1985;39(2-3):155-63
PMID: 4092928
-
Conserved structural features are found upstream from the three co-ordinately regulated discoidin I genes of Dictyostelium discoideum.
J Mol Biol. 1984 Jan 15;172(2):203-20
PMID: 6694210
-
Overexpression of the csA cell adhesion molecule under its own cAMP-regulated promoter impairs morphogenesis in Dictyostelium.
J Cell Sci. 1992 Jun;102 ( Pt 2):203-14
PMID: 1328260
-
Establishment of a system for conditional gene expression using an inducible tRNA suppressor gene.
Mol Cell Biol. 1992 Sep;12(9):4038-45
PMID: 1508201
-
A cis-acting element responsible for early gene induction by extracellular cAMP in Dictyostelium discoideum.
Mech Dev. 1991 Feb;33(2):147-55
PMID: 1851628
-
Constitutive overexpression of the contact site A glycoprotein enables growth-phase cells of Dictyostelium discoideum to aggregate.
EMBO J. 1990 Sep;9(9):2709-16
PMID: 2390970
-
Identification of a cis-acting element controlling induction of early gene expression in Dictyostelium discoideum.
Mol Cell Biol. 1989 Nov;9(11):4653-9
PMID: 2601692
-
Developmental regulation of Dictyostelium discoideum actin gene fusions carried on low-copy and high-copy transformation vectors.
Mol Cell Biol. 1986 Nov;6(11):3973-83
PMID: 3025622
-
Antisense RNA inactivation of myosin heavy chain gene expression in Dictyostelium discoideum.
Science. 1987 May 29;236(4805):1081-6
PMID: 3576221
-
Structural and functional characterization of a Dictyostelium gene encoding a DIF inducible, prestalk-enriched mRNA sequence.
Nucleic Acids Res. 1987 Sep 25;15(18):7463-76
PMID: 3658700
-
Differential expression of the members of the discoidin I multigene family during growth and development of Dictyostelium discoideum.
Cell. 1982 Apr;28(4):793-800
PMID: 6284373
-
Expression of human antithrombin III in the cellular slime mould Dictyostelium discoideum.
Appl Microbiol Biotechnol. 1991 Jul;35(4):496-503
PMID: 1367393
-
Dictyostelium discoideum as an expression host for the circumsporozoite protein of Plasmodium falciparum.
Gene. 1992 Feb 15;111(2):157-63
PMID: 1541397
-
Overproduction of the regulatory subunit of the cAMP-dependent protein kinase blocks the differentiation of Dictyostelium discoideum.
EMBO J. 1989 Jul;8(7):2039-43
PMID: 2551673