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DNA methylases separated through the HeLa cell cycle methodology show allosteric properties
Author(s) -
Delfini C.,
Crema A.L.,
Alfani E.,
Lo Presti E.,
Eremenko T.,
Volpe P.
Publication year - 1987
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(87)81289-9
Subject(s) - hpaii , pbr322 , plasmid , chemistry , restriction enzyme , dna , allosteric regulation , microbiology and biotechnology , hela , biology , biochemistry , enzyme , gene , cell , dna methylation , gene expression
Two DNA methylases (DNAmets) can be separated through the cell cycle. The first appears as a minor peak in G 1 , the second as a major peak in S. Both enzymes protect from Hpa ll a plasmid (H 31 ), constructed with the pBR322 vector (4.3 kbp) and the inverted A γ , fragment of the human globin gene (3.5 kbp), inserted at its Hind III site (the vector carries several Hpa II sites, the insert only one Hpa II site). DNAmets G 1 and S show distinct K m values and different kinetics vs the ionic strength of the medium, while their Michaelis‐Menten and Lineweaver‐Burk plots are sigmoidal and hyperbolical curves, respectively. This is the first suggestion about the allosteric nature of the eukaryotic DNAmet system.