Abstract:
:We have developed a cell-free system for studying the integration of retroviral DNA. In our assay, amber mutations in a bacteriophage lambda genome that serves as the target for integration are suppressed by integration of an MLV derivative that carries the E. coli supF gene. The structure of the reaction products is that expected from an authentic MLV integration reaction. Linear viral DNA from the cytoplasm of infected cells serves as a precursor, though not necessarily the immediate precursor, to the provirus integrated in vitro. The viral DNA in the infected cell appears to be tightly associated with the enzymatic machinery required for its integration. Supercoiling, chromatin structure, transcription, and replication are not required of the target DNA. Since no high-energy cofactor is necessary, the DNA breakage and joining steps in the integration reaction are probably coupled.
journal_name
Celljournal_title
Cellauthors
Brown PO,Bowerman B,Varmus HE,Bishop JMdoi
10.1016/0092-8674(87)90287-xsubject
Has Abstractpub_date
1987-05-08 00:00:00pages
347-56issue
3eissn
0092-8674issn
1097-4172pii
0092-8674(87)90287-Xjournal_volume
49pub_type
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