Identification of the Streptococcus mutans frp gene as a potential regulator of fructosyltransferase expression.
Four putative open reading frames (ORFs) were previously identified in the regions flanking the Streptococcus mutans GS-5 fructosyltransferase (FTF) gene. One of these, ORF 3, appeared to code for a low-molecular-mass protein containing amino acid sequences sharing homology with several Gram-positive bacterial DNA-binding proteins and it was suggested that the ORF 3 gene product might be an FTF regulatory protein (FRP). In order to characterize this protein, we have purified the biotinylated tag-FRP fusion protein using the PinPoint protein purification system and this fusion protein was used in gel shift assays with DNA fragments containing the ftf promoter region. FRP bound specifically to the upstream region of the ftf promoter containing the inverted repeat structure that is present upstream of the -35 sequence. In contrast, FRP did not bind to DNA fragments lacking the inverted repeat structure. The results of these experiments suggest that FRP interacts with the inverted repeat region upstream of the ftf promoter and such interactions may regulate FTF expression.
Shibata
Y
Y
Department of Oral Biology, School of Dental Medicine, State University of New York at Buffalo 14214-3092, USA.
Kuramitsu
H K
HK
D000595:Amino Acid Sequence; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D038702:Follistatin-Related Proteins; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D005798:Genes, Bacterial / Q000235:genetics; D006023:Glycoproteins / Q000235:genetics* / Q000302:isolation & purification; D006602:Hexosyltransferases / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D013295:Streptococcus mutans / Q000201:enzymology / Q000235:genetics*; D001426:Bacterial Proteins
D002352:Carrier Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D038702:Follistatin-Related Proteins; D006023:Glycoproteins; D006602:Hexosyltransferases; C021451:inulosucrase; D001426:Bacterial Proteins