DNA binding protein from Streptomyces hygroscopicus: detection of binding by gel retardation, sedimentation and effects on the transcriptional activity in vitro.
The binding to supercoiled plasmid DNA as well as to double-stranded short DNA fragments of the DNA binding protein. ASBIII, from S. hygroscopicus has been demonstrated by gel retardation asay. As further revealed by sedimentation analysis, the protein-DNA complex formation also involves condensation of DNA and an aggregation step at higher total protein-to-DNA ratio (w/w). In vitro studies of the transcriptional activity of DNA-protein complexes showed that the ASBIII protein inhibits the overall template activity in the RNA polymerase II system to nearly similar extents for various DNA's. Preincubation of ASBIII-DNA complexes of different GC content, however, suggested a selective inhibitory effect on GC-rich DNA due to the higher stability of this complex. The results also indicate that the initiation of transcription of the GC-rich DNA template is affected by the ASBIII protein. The present results on the DNA binding properties and inactivation of the transcription of the ASBIII protein suggest that this protein may be a potential candidate for the local compaction of chromosomal DNA in Streptomyces hygroscopicus.
Sarfert
E
E
Academy of Sciences of GDR, Department of Molecular Biochemistry.
Sedova
V
V
Triebel
H
H
Bär
H
H
Zimmer
C
C
D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D002850:Chromatography, Gel; D004262:DNA Restriction Enzymes; D004268:DNA-Binding Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D066298:In Vitro Techniques; D008099:Liver / Q000378:metabolism; D010761:Phosphorus Radioisotopes; D010957:Plasmids; D012319:RNA Polymerase II / Q000378:metabolism; D051381:Rats; D013302:Streptomyces / Q000378:metabolism*; D014158:Transcription, Genetic; D014461:Ultracentrifugation
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D010761:Phosphorus Radioisotopes; D012319:RNA Polymerase II; D004262:DNA Restriction Enzymes