title: Identification of conserved features of LAGLIDADG homing endonucleases.
abstract: LAGLIDADG family of homing endonucleases are rare-cutting enzymes which recognize long target sequences and are of great interest in genome engineering. Despite advances in homing endonuclease engineering, effective methods of broadening the range of cleaved sequences are still lacking. Here, we present a study of conserved structural features of LAGLIDADG homing endonucleases that might aid further development of such methods. The protein-DNA interface of LAGLIDADG homing endonucleases differs considerably with the particular nuclease, and the analysis of conserved protein-DNA interactions could not identify any residues crucial for DNA binding and common to most nucleases of the family. For the homing endonuclease PI-SceI, a comparison of structural and experimental data derived from literature helped to identify 23 residues that are likely to be important for DNA binding. Analysis of the LAGLIDADG domain dimerization interface allowed the choosing of six positions that contribute to dimerization specificity most, while comparison of 446 sequences of LAGLIDADG endonucleases revealed groups of residues in these positions that appear to be most favorable for dimerization.
authors.affiliation: AN Belozersky Institute for Physical and Chemical Biology, Moscow State University, Moscow 119991, Russia. grishin-a1@yandex.ru
authors.forename: Alexander
authors.initials: A
authors.lastname: Grishin
authors.forename: Ines
authors.initials: I
authors.lastname: Fonfara
authors.forename: Andrei
authors.initials: A
authors.lastname: Alexeevski
authors.forename: Sergei
authors.initials: S
authors.lastname: Spirin
authors.forename: Olga
authors.initials: O
authors.lastname: Zanegina
authors.forename: Anna
authors.initials: A
authors.lastname: Karyagina
authors.forename: Daniil
authors.initials: D
authors.lastname: Alexeyevsky
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Wende
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D004247:DNA / Q000737:chemistry* / Q000235:genetics*; D003850:Deoxyribonuclease I / Q000737:chemistry* / Q000235:genetics*; D008969:Molecular Sequence Data; D011485:Protein Binding; D017422:Sequence Analysis, DNA / Q000379:methods*; D020539:Sequence Analysis, Protein / Q000379:methods*
chemical_list: D004247:DNA; D003850:Deoxyribonuclease I