Cloning and characterization of the G-box binding factor, an essential component of the developmental switch between early and late development in Dictyostelium.
During Dictyostelium development, the cAMP-regulated induction of cell-type-specific late genes marks a developmental switch from the initial formation of the multicellular organism to the differentiation of the various cell types that mediate morphogenesis and eventually give rise to the mature fruting body. The G-box binding factor (GBF) is a developmentally regulated Dictyostelium transcription factor whose affinity for a DNA sequence correlates with the ability of that sequence to confer inducibility to late gene promoters in response to high, continuous levels of extracellular cAMP. We report the purification of GBF and cloning of the gene that encodes it, as confirmed by in vitro production of GBF activity. The predicted protein is highly basic and contains two putative zinc fingers. Disruption of the GBF gene by homologous recombination results in the loss of all GBF DNA-binding activity, developmental arrest at the loose aggregate stage, and the loss of late gene induction during development or in response to extracellular cAMP. Constitutive expression of GBF complements the null phenotype and allows for the rapid activation of a class of late genes in response to cAMP. Our results indicate that GBF acts as an extracellular cAMP-responsive transcriptional activator regulating late gene expression and is an essential component of a developmental switch between aggregation and cellular morphogenesis.
Schnitzler
G R
GR
Department of Biology, University of California, San Diego, La Jolla 92093-0634.
Fischer
W H
WH
Firtel
R A
RA
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D000242:Cyclic AMP / Q000494:pharmacology; D004271:DNA, Fungal / Q000235:genetics; D016054:DNA, Protozoan / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000502:physiology; D004023:Dictyostelium / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development*; D005656:Fungal Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000502:physiology; D051790:G-Box Binding Factors; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D005800:Genes, Fungal / Q000187:drug effects; D017125:Genes, Protozoan / Q000187:drug effects; D008969:Molecular Sequence Data; D008970:Molecular Weight; D015800:Protozoan Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D014157:Transcription Factors; D015533:Transcriptional Activation; D016335:Zinc Fingers / Q000235:genetics
D004271:DNA, Fungal; D016054:DNA, Protozoan; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D051790:G-Box Binding Factors; D015800:Protozoan Proteins; D014157:Transcription Factors; D000242:Cyclic AMP