Abstract
Strains of Neisseria gonorrhoeae possess numerous restriction-modification (R-M) systems. One of these systems, which has been found in all strains tested, encodes the S. NgoVIII specificity (5'TCACC 3') R-M system. We cloned two adjacent methyltransferase genes (dcmH and damH), each encoding proteins whose actions protect DNA from digestion by R.HphI or R.Ngo BI (5'TCACC 3'). The damH gene product is a N 6-methyladenine methyltransferase that recognizes this sequence. We constructed a plasmid containing multiple copies of the S.NgoVIII sequence, grew it in the presence of damH and used the HPLC to demonstrate the presence of N 6-methyladenine in the DNA. A second plasmid, containing overlapping damH and Escherichia coli dam recognition sequences in combination with various restriction digests, was used to identify which adenine in the recognition sequence was modified by damH. The predicted dcmH gene product is homologous to 5-methylcytosine methyltransferases. The products of both the dcmH and damH genes, as well as an open reading frame downstream of the damH gene are highly similar to the Haemophilus parahaemolyticus hphIMC , hphIMA and hphIR gene products, encoding the Hph I Type IIs R-M system. The S.NgoVIII R-M genes are flanked by a 97 bp direct repeat that may be involved in the mobility of this R-M system.
MeSH Terms
Amino Acid Sequence
Cloning, Molecular
DNA Restriction-Modification Enzymes/genetics
DNA, Bacterial/chemistry,genetics
DNA-Cytosine Methylases/chemistry,genetics
Haemophilus/enzymology,genetics
Molecular Sequence Data
Neisseria gonorrhoeae/enzymology,genetics
Open Reading Frames
Restriction Mapping
Sequence Analysis, DNA
Sequence Homology
Chemicals
DNA Restriction-Modification Enzymes
DNA, Bacterial
DNA modification methylase NgoBI
DNA-Cytosine Methylases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Gunn J S
University of Maryland, Department of Microbiology, College Park, MD 20142, USA.
Stein D C
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