DNA-binding specificity of the CYS3 transcription factor of Neurospora crassa defined by binding-site selection.
The CYS3 transcription factor is a basic region-leucine zipper (bZIP) DNA-binding protein that is essential for the expression of a coordinately regulated group of genes involved in the acquisition and utilization of sulfur in Neurospora crassa. An approach of using binding-site selection from random-sequence oligonucleotides was used to define CYS3-binding specificity. The derived consensus-binding site of ATGGCGCCAT defines a symmetrical sequence (half-site A T G/t G/a C/t) that resembles that of other bZIP proteins such as CREB and C/EBP. By comparison, CYS3 shows a greater range of binding to a central core of varied Pur-Pyr-Pur-Pyr sequences than CREB as determined by gel shift assays. The derived CYS3 consensus binding sequence was further validated by demonstrating in vivo sulfur regulation using a heterologous promoter construct. The CYS3-binding site data will be useful for the genome-wide study of sulfur-regulated genes in N. crassa, which has served as a model fungal sulfur control system.
Paietta
John V
JV
Department of Biochemistry and Molecular Biology, Wright State University, Dayton, OH 45435, USA. john.paietta@wright.edu
D001483:Base Sequence; D050976:Basic-Leucine Zipper Transcription Factors / Q000378:metabolism; D001665:Binding Sites / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D005656:Fungal Proteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D009492:Neurospora crassa / Q000378:metabolism*; D009841:Oligonucleotides / Q000737:chemistry / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011485:Protein Binding; D014157:Transcription Factors / Q000378:metabolism*
D050976:Basic-Leucine Zipper Transcription Factors; D005656:Fungal Proteins; D009841:Oligonucleotides; D014157:Transcription Factors