title: In vitro murine leukemia retroviral integration and structure fluctuation of target DNA.
abstract: Integration of the retroviral genome into host DNA is a critical step in the life cycle of a retrovirus. Although assays for in vitro integration have been developed, the actual DNA sequences targeted by murine leukemia retrovirus (MLV) during in vitro reproduction are unknown. While previous studies used artificial target sequences, we developed an assay using target DNA sequences from common MLV integration sites in Stat5a and c-myc in the genome of murine lymphomas and successfully integrated MLV into the target DNA in vitro. We calculated the free energy change during folding of the target sequence DNA and found a close correlation between the calculated free energy change and the number of integrations. Indeed, the integrations closely correlated with fluctuation of the structure of the target DNA segment. These data suggest that the fluctuation may generate a DNA structure favorable for in vitro integration into the target DNA. The approach described here can provide data on the biochemical properties of the integration reaction to which the target DNA structure may contribute.
authors.affiliation: Department of Forensic Medicine and Molecular Pathology, Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto, Japan. tsuruyam@kuhp.kyoto-u.ac.jp
authors.forename: Tatsuaki
authors.initials: T
authors.lastname: Tsuruyama
authors.forename: Weizhi
authors.initials: W
authors.lastname: Liu
authors.forename: Kenichi
authors.initials: K
authors.lastname: Yoshikawa
mesh_terms: D000818:Animals; D001483:Base Sequence; D004279:DNA, Viral / Q000737:chemistry*; D018390:Gene Targeting; D016679:Genome, Viral; D009052:Leukemia Virus, Murine / Q000235:genetics*; D051379:Mice; D009690:Nucleic Acid Conformation; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics; D012190:Retroviridae / Q000235:genetics; D050799:STAT5 Transcription Factor / Q000235:genetics; D013816:Thermodynamics; D016662:Virus Integration
chemical_list: D004279:DNA, Viral; C489428:Myc protein, mouse; D016271:Proto-Oncogene Proteins c-myc; D050799:STAT5 Transcription Factor; C494093:Stat5a protein, mouse