Proteins encoded by the DpnII restriction gene cassette. Two methylases and an endonuclease.
Proteins encoded by three genes in the DpnII restriction enzyme cassette of Streptococcus pneumoniae were purified and characterized. Large amounts of the proteins were produced by subcloning the cassette in an Escherichia coli expression system. All three proteins appear to be dimers composed of identical polypeptide subunits. One is the DpnII endonuclease, and the other two are DNA adenine methylase active at 5' GATC 3' sites. Inactivation of enzyme activity by insertions into the genes and comparison of the DNA sequence with the amino-terminal sequence of amino acid residues in the proteins demonstrated the following correspondence between genes and enzymes. The promoter-proximal gene in the operon, dpnM, encodes a 33 X 10(3) Mr polypeptide that gives rise to a potent DNA methylase. The next gene, dpnA, encodes the 31 x 10(3) Mr polypeptide of a weaker and less-specific methylase. The third gene, dpnB, encodes the 34 x 10(3) Mr polypeptide of the endonuclease. Although the endonuclease polypeptide is initiated from an ordinary ribosome-binding site, each of the methylase polypeptide begins at an atypical site with a consensus sequence entirely different from that of Shine & Dalgarno. This presumptive novel ribosome-binding site is well recognized in both S. pneumoniae and E. coli.
de la Campa
A G
AG
Department of Biology, Brookhaven National Laboratory, Upton, NY 11973.
Kale
P
P
Springhorn
S S
SS
Lacks
S A
SA
D000595:Amino Acid Sequence; D002850:Chromatography, Gel; D004262:DNA Restriction Enzymes / Q000235:genetics* / Q000302:isolation & purification; D015252:Deoxyribonucleases, Type II Site-Specific; D005798:Genes, Bacterial; D008780:Methyltransferases / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D008970:Molecular Weight; D014176:Protein Biosynthesis; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific); D013296:Streptococcus pneumoniae / Q000201:enzymology
D008780:Methyltransferases; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific); D004262:DNA Restriction Enzymes; D015252:Deoxyribonucleases, Type II Site-Specific; C024405:GATC-specific type II deoxyribonucleases