title: Restriction endonuclease AgeI is a monomer which dimerizes to cleave DNA.
abstract: Although all Type II restriction endonucleases catalyze phosphodiester bond hydrolysis within or close to their DNA target sites, they form different oligomeric assemblies ranging from monomers, dimers, tetramers to higher order oligomers to generate a double strand break in DNA. Type IIP restriction endonuclease AgeI recognizes a palindromic sequence 5΄-A/CCGGT-3΄ and cuts it ('/' denotes the cleavage site) producing staggered DNA ends. Here, we present crystal structures of AgeI in apo and DNA-bound forms. The structure of AgeI is similar to the restriction enzymes that share in their target sites a conserved CCGG tetranucleotide and a cleavage pattern. Structure analysis and biochemical data indicate, that AgeI is a monomer in the apo-form both in the crystal and in solution, however, it binds and cleaves the palindromic target site as a dimer. DNA cleavage mechanism of AgeI is novel among Type IIP restriction endonucleases.
authors.affiliation: Institute of Biotechnology, Vilnius University, Sauletekio al. 7, LT-10257 Vilnius, Lithuania.
authors.forename: Giedre
authors.initials: G
authors.lastname: Tamulaitiene
authors.affiliation: Institute of Biotechnology, Vilnius University, Sauletekio al. 7, LT-10257 Vilnius, Lithuania.
authors.forename: Virginija
authors.initials: V
authors.lastname: Jovaisaite
authors.affiliation: Institute of Biotechnology, Vilnius University, Sauletekio al. 7, LT-10257 Vilnius, Lithuania.
authors.forename: Gintautas
authors.initials: G
authors.lastname: Tamulaitis
authors.affiliation: Institute of Biotechnology, Vilnius University, Sauletekio al. 7, LT-10257 Vilnius, Lithuania.
authors.forename: Inga
authors.initials: I
authors.lastname: Songailiene
authors.affiliation: Institute of Biotechnology, Vilnius University, Sauletekio al. 7, LT-10257 Vilnius, Lithuania.
authors.forename: Elena
authors.initials: E
authors.lastname: Manakova
authors.affiliation: Institute of Biotechnology, Vilnius University, Sauletekio al. 7, LT-10257 Vilnius, Lithuania.
authors.forename: Mindaugas
authors.initials: M
authors.lastname: Zaremba
authors.affiliation: Institute of Biotechnology, Vilnius University, Sauletekio al. 7, LT-10257 Vilnius, Lithuania.
authors.forename: Saulius
authors.initials: S
authors.lastname: Grazulis
authors.affiliation: New England Biolabs, Inc. 240 County Road, Ipswich, MA 01938, USA.
authors.forename: Shuang-Yong
authors.initials: SY
authors.lastname: Xu
authors.affiliation: Institute of Biotechnology, Vilnius University, Sauletekio al. 7, LT-10257 Vilnius, Lithuania.
authors.forename: Virginijus
authors.initials: V
authors.lastname: Siksnys
mesh_terms: D001051:Apoenzymes / Q000737:chemistry; D020029:Base Pairing; D020134:Catalytic Domain; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D053837:DNA Cleavage; D015252:Deoxyribonucleases, Type II Site-Specific / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D011485:Protein Binding; D055503:Protein Multimerization
chemical_list: D001051:Apoenzymes; D004247:DNA; D015252:Deoxyribonucleases, Type II Site-Specific