Open AccessIdentification of the Streptococcus mutans frp gene as a potential regulator of fructosyltransferase expression
Author(s) -
Shibata Yukie,
Kuramitsu Howard K
Publication year - 1996
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1996.tb08313.x
Subject(s) - orfs , biology , open reading frame , gene , dna , biotinylation , inverted repeat , fusion protein , microbiology and biotechnology , genetics , peptide sequence , recombinant dna , genome
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 fif 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.