Phased A-tracts bind to the alpha subunit of RNA polymerase with increased affinity at low temperature.
Previously we showed that the expression of a Clostridium perfringens phospholipase C gene (plc) is activated by promoter upstream phased A-tracts in a low temperature-dependent manner. In this paper we characterize the interaction between the alpha subunit of C. perfringens RNA polymerase and the phased A-tracts. Hydroxyl radical footprinting and fluorescence polarization assaying revealed that the alpha subunit binds to the minor grooves of the phased A-tracts through its C-terminal domain with increased affinity at low temperature. The result provides a molecular mechanism underlying the activation of the plc promoter by the phased A-tracts.
Katayama
S
S
Department of Biochemistry and Chemistry, Faculty of Science, Okayama University of Science, 1-1, Ridai-cho, Okayama 700-0005, Japan.
Matsushita
O
O
Tamai
E
E
Miyata
S
S
Okabe
A
A
D001483:Base Sequence; D001665:Binding Sites; D003016:Clostridium perfringens; D004269:DNA, Bacterial / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D021122:Protein Subunits; D010738:Type C Phospholipases / Q000235:genetics*
D004269:DNA, Bacterial; D021122:Protein Subunits; D012321:DNA-Directed RNA Polymerases; C113275:RNA polymerase alpha subunit; D010738:Type C Phospholipases