Determination of the recognition site for adenine-specific methylase of Shigella sonnei 47 by hydazinolysis of DNA, followed by separation of the purine oligonucleotides by thin-layer chromatography on DEAE-cellulose

Journal Article · Fri Sep 20 00:00:00 EDT 1985 · Biochemistry (Engl. Transl.); (United States)

OSTI ID:5722359

Lopatina, N G; Kirnos, M D; Suchkov, S V; Vanyushin, B F; Nikol'skaya, I I; Debov, S S

A method has been developed for the separation of oligopurine units according to length and composition by two-dimensional thin-layer chromatography on plates with DEAE-cellulose, permitting a comparative analysis of the content of various purine isopliths in DNA of different origin. In the case of the analysis of methylated DNA, the method permits a comparison of the substrate specificity of various enzymes of methylation of the adenine residues in DNA. In conjunction with enzymatic treatment of labeled methylated isopliths, the method permits determination of the methylatable sequence and in a number of cases an ascertainment of the recognition site for adenine-specific methylase as a whole. The proposed method was used to establish the fact that the methylase Ssol recognizes the sequence 5'...G-A-A-T-T-C...3' and methylates the adenine residue closest to its 5'-end.

• Research Organization: Scientific-Research Institute of Medical Enzymology, Moscow, USSR
• OSTI ID: 5722359
• Journal Information: Biochemistry (Engl. Transl.); (United States), Vol. 50:3; Other Information: Translation from Biokhimiya; 50: No. 3, 495-502(Mar 1985)
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
ADENINES
METHYLATION
RADIOENZYMATIC ASSAY
RADIORECEPTOR ASSAY
DNA
ENZYMATIC HYDROLYSIS
NUCLEOTIDES
THIN-LAYER CHROMATOGRAPHY
PURINES
LABELLING
RADIONUCLIDE KINETICS
RECEPTORS
SHIGELLA
TRITIUM COMPOUNDS
UPTAKE
AMINES
ANTIMETABOLITES
AROMATICS
AZAARENES
BACTERIA
CHEMICAL REACTIONS
CHROMATOGRAPHY
DECOMPOSITION
DRUGS
HETEROCYCLIC COMPOUNDS
HYDROLYSIS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LYSIS
MEMBRANE PROTEINS
MICROORGANISMS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PROTEINS
SEPARATION PROCESSES
SOLVOLYSIS
TRACER TECHNIQUES
550201* - Biochemistry- Tracer Techniques