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PMID:17188297
| Citation |
Bair, CL and Black, LW (2007) A type IV modification dependent restriction nuclease that targets glucosylated hydroxymethyl cytosine modified DNAs. J. Mol. Biol. 366:768-78 |
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| Abstract |
The Escherichia coli CT596 prophage exclusion genes gmrS and gmrD were found to encode a novel type IV modification-dependent restriction nuclease that targets and digests glucosylated (glc)-hydroxymethylcytosine (HMC) DNAs. The protein products GmrS (36 kDa) and GmrD (27 kDa) were purified and found to be inactive separately, but together degraded several different glc-HMC modified DNAs (T4, T2 and T6). The GMR enzyme is able to degrade both alpha-glucosy-HMC T4 DNA and beta-glucosyl-HMC T4 DNA, whereas no activity was observed against non-modified DNAs including unmodified T4 cytosine (C) DNA or non-glucosylated T4 HMC DNA. Enzyme activity requires NTP, favors UTP, is stimulated by calcium, and initially produces 4 kb DNA fragments that are further degraded to low molecular mass products. The enzyme is inhibited by the T4 phage internal protein I* (IPI*) to which it was found to bind. Overall activities of the purified GmrSD enzyme are in good agreement with the properties of the cloned gmr genes in vivo and suggest a restriction enzyme specific for sugar modified HMC DNAs. IPI* thus represents a third generation bacteriophage defense against restriction nucleases of the Gmr type. |
| Links |
PubMed PMC1855630 Online version:10.1016/j.jmb.2006.11.051 |
| Keywords |
Capsid Proteins/metabolism; Chromatography, Affinity; Coliphages; Cytosine/analogs & derivatives; Cytosine/metabolism; DNA Restriction Enzymes/metabolism; DNA, Viral/chemistry; DNA, Viral/metabolism; Escherichia coli/enzymology; Escherichia coli/virology; Escherichia coli Proteins/isolation & purification; Escherichia coli Proteins/metabolism; Evolution, Molecular; Mutant Proteins/metabolism; Myoviridae; Nucleic Acid Conformation; Nucleotides/metabolism; Protein Binding; Site-Specific DNA-Methyltransferase (Adenine-Specific)/metabolism; Substrate Specificity |
| edit table |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0032067: Type IV site-specific deoxyribonuclease activity |
ECO:0000315: |
F |
Figure 2, Figure 4. IPI protects wtT4 DNA, from GmrSD that digest DNAs. Agarose gel stained with ethidium bromide showed the activity of wtT4 with IPI. |
complete | ||||
|
enables |
GO:0032067: type IV site-specific deoxyribonuclease activity |
ECO:0000315: mutant phenotype evidence used in manual assertion |
F |
Seeded From UniProt |
complete | |||
See also
References
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