Klimasauskas, S., Nelson, J. L., Roberts, R. J. (November 1991) The sequence specificity domain of cytosine-C5 methylases. Nucleic Acids Res, 19 (22). pp. 6183-90. ISSN 0305-1048 (Print)0305-1048 (Linking)
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Abstract
Prokaryotic DNA[cytosine-C5]methyltransferases (m5C-methylases) share a common architectural arrangement of ten conserved sequence motifs. A series of eleven hybrids have been constructed between the HpaII (recognition sequence: Cm5CGG) and HhaI (recognition sequence: Gm5CGC) DNA-methylases. The hybrids were over-expressed in E.coli and their in vivo methylation phenotypes investigated. Six were inactive by our assay while five of them retained partial methylation activity and full specificity. In all five cases the specificity matched that of the parent methylase which contributed the so-called variable region, located between conserved motifs VIII and IX. This was the only sequence held in common between the active hybrids and for the first time provides unequivocal evidence that the specificity determinants of the mono-specific m5C-methylases are located within the variable region. Correlation of the hybrid methylase structure with the efficiency of methylation suggests that conserved motif IX may interact with the variable region whereas motif X most probably interacts with the N-terminal half of the molecule.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | Amino Acid Sequence Base Sequence DNA Restriction Enzymes DNA, Bacterial/genetics DNA-Cytosine Methylases/*genetics Escherichia coli/enzymology/genetics Genes, Bacterial Methylation Molecular Sequence Data Mutagenesis Plasmids Polymerase Chain Reaction Sequence Alignment |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > methyltransferase bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > restriction enzyme |
| CSHL Authors: | |
| Communities: | CSHL labs > Roberts lab |
| Depositing User: | Matt Covey |
| Date: | 25 November 1991 |
| Date Deposited: | 10 Dec 2015 16:49 |
| Last Modified: | 08 Nov 2017 16:42 |
| PMCID: | PMC329119 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/32142 |
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