DNA cleavage by restriction endonuclease PflMI is inhibited in recognition sites modified by dcm methylation

Sturm, R. A., Yaciuk, P. (1989) DNA cleavage by restriction endonuclease PflMI is inhibited in recognition sites modified by dcm methylation. Nucleic Acids Res, 17 (9). p. 3615. ISSN 0305-1048

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URL: http://www.ncbi.nlm.nih.gov/pubmed/2657665

Abstract

Plasmids pBSoc£-l + (1) and pALTEla (described below*) both contain two canonical P/IMI recognition sites [CCANNNNNTGG (2)]. These plasmids, when purified from Escherichia coli dam/dcm~ strain RB404 (3). release a 362- or 337-bp fragment, respectively, after digestion with P/IMI (Fig. 1A, lanes 1 and 6). However, when purified from dam*/dcm* E. coli strains, these plasmids cleaved only once after digestion with P/IMI (Fig. 1A, lanes 2 and 7). Digestion of unmethylated (lane 3) and methylated (lane 4) pBSocM* DNA with P/IMI and EcoRI Identified the oct-1 P/IMI site at nucleotide position 1680 (1) (see Fig. IB) as the recognition site that is resistant to cleavage. Digestion of unmethylated (lane 8) and methylated (lane 9) pALTEla DNA with P/IMI and PuuII identified the adenovirus P/IMI site at nucleotide position 1461 (4) (see Fig 1C) as the resistant site. Both cleavage-resistant P/IMI sites contain the dcm methylation sequence, CmCWGG (5). These results suggest that a subset of P/IMI recognition sequences that contain a dcm methylation site are resistant to P/IMI cleavage when methylated.

Item Type: Paper
Uncontrolled Keywords: Base Sequence DNA, Bacterial/genetics/metabolism Deoxyribonucleases, Type II Site-Specific/*metabolism Escherichia coli/genetics Molecular Sequence Data *Plasmids Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Restriction Mapping
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > DNA methylation
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > restriction enzyme
CSHL Authors:
Communities: CSHL labs
Depositing User: Gail Sherman
Date Deposited: 23 Jun 2017 19:51
Last Modified: 08 Nov 2017 16:59
PMCID: PMC317826
Related URLs:
URI: http://repository.cshl.edu/id/eprint/34911

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